Vegetable oil composition containing palm mid-fraction fat and method of reducing plasma cholesterol

ABSTRACT

A method and composition for reducing the cholesterolemic effect in mammals of ingesting a blended nutritional fat composition containing a palm mid-fraction (PMF) hardstock fat combined with an unsaturated vegetable oil. The composition is solid or semi-solid at 20° C. and fluid at 35° C., and includes between 15% and 45% by weight linoleic acid. The weight ratio of disaturated triglyceride (DST) molecules to trisaturated triglyceride (TST) molecules is greater than 10:1, and the PMF hardstock fat contains approximately 50% to 95% by weight DST molecules, the majority of which contain either palmitic acid or a combination of palmitic and stearic acids at the sn-1 and sn-3 triglyceride positions and either oleic acid or linoleic acid at the sn-2 molecular position.

BACKGROUND

Previous clinical research has described dietary fats and their role inmodulating major species of plasma lipoproteins (Mensink et al. 2003; AmJ Clin Nutr, 77:1146-1155), as well as their role in coronary heartdisease and controlling plasma cholesterol levels (Steinberg et al.1999; JAMA, 282(21): 2043-2050). Other research has studied changes inlipoprotein levels resulting from dietary fats that are rich in variousfatty acids. For example, Tholstrup et al. (1994; Am J Clin Nutr,59:371-377) studied changes in lipoprotein levels resulting from dietsrich in different saturated fatty acids including stearic acid, palmiticacid, and lauric and myristic acids. Researchers have also studied andcompared the abilities of different fatty acids to raise or loweroverall cholesterol levels in human plasma. Most nutritional expertsagree that saturated fatty acids as a class raise total cholesterollevels, while polyunsaturated fatty acids lower them. Monounsaturatedfatty acids, e.g., oleic acid, are considered more neutral in theireffect. It is also understood that the metabolism of individual fattyacid species within each class can impact HDL and LDL cholesterol levelsto different degrees.

A number of research studies have suggested that, of all the more commonsaturated fatty acids, including lauric acid (C12:0), myristic acid(C14:0), palmitic acid (C16:0), and stearic acid (C18:0), it is myristicacid that is the most potent in elevating total cholesterol levels inplasma. Consistent with these findings, some manufacturers of processedfoods avoid the use of hardening fats such as coconut oil or palm kerneloil, which contain high levels of myristic acid, in favor of palmstearin and regular palm oil, which are also hardening fats but containhigh levels of palmitic and stearic acids instead.

A recently produced commercial margarine known as SMART BALANCE butteryspread (GFA Brands, Inc., Paramus, N.J.) combines the beneficial LDLcholesterol-lowering properties of polyunsaturated fatty acids, e.g.,found in soybean oil, with the beneficial HDL cholesterol-raising andoil hardening properties of saturated fats. SMART BALANCE margarineincorporates regular palm oil which is rich in palmitic acid, ratherthan palm kernel oil which is rich in lauric and myristic acids, toachieve the requisite hardened texture. This margarine and relatedhealthful fat blends are based upon the work of Sundram et al., and isdescribed in U.S. Pat. Nos. 5,578,334, 5,843,497, 6,630,192 and7,229,653, which are incorporated herein in their entireties. Sundram etal. describe a cholesterol-free blended fat composition that combines apolyunsaturated fat (15-40 wt % linoleic acid), and a cholesterol-freesaturated fat in which the saturated fatty acids provide between 20% and40% by weight of the composition. The effect of the saturated fat, i.e.,palm oil, in this margarine is to increase both HDL and LDL cholesterolwhile the effect of the polyunsaturated vegetable oil is to lower LDLcholesterol. The net effect of periodically or regularly consuming sucha fat blend composition instead of typical American dietary fat wasshown to be a modest increase in the HDL concentration and an increasein the HDL/LDL concentration ratio in the blood.

Subfractions of palm oil, including palm stearin and so-called palmmid-fractions, have been commercially prepared without any chemicalmodification and used as hardstocks to solidify vegetable oils inmargarines and table spreads. The palm stearins are differentiated fromthe mid-fractions in that stearins contain a high level of trisaturatedtriglycerides (e.g., tripalmitin or PPP) resulting in an elevatedmelting point (typically about 54° C.) whereas palm mid-fractionscontain a significantly reduced level of PPP and an elevated level ofdisaturated triglycerides (e.g., POP containing two palmitates and oneoleate fatty acid) resulting in a beneficially reduced melting point(about 32° C.). Commercial stearins and mid-fractions are available withfairly similar iodine values (IV level, a measure of the amount ofunsaturation in fat; grams of iodine consumed by 100 grams of fat) thatare compatible with margarine and shortening use (e.g., with medium IVlevels of 30-35). The use of a similar IV level palm stearin and palmmid-fraction can provide similar texture or “softness” in the fat,suitable for margarine and shortening use.

Patents related to the use of palm mid-fraction in margarines andspreads include U.S. Pat. No. 4,115,598 (Moran), U.S. Pat. No. 4,388,339(Lomneth), U.S. Pat. No. 4,390,561 (Blair), and U.S. Pat. No. 4,568,556(McCoy). These patents describe the use of so-called palm mid-fractionsas structural fats for solidifying vegetable oils such as soybean oiland sunflower oil. The functionally important solid fat content measuredat room temperature for different preparations of palm mid-fraction canvary widely depending on the content of trisaturated, disaturated, andmonosaturated triglycerides. Moran describes oil-in-water emulsionshaving 60% aqueous phase and 40% oil phase, in which the oil phasecontains high levels of a palm mid-fraction (25%-30% by weight).However, such high levels of saturated fatty acids tend to elevate totalcholesterol levels in human plasma. The palm mid-fraction of Moran iscombined with either high levels of sunflower oil (70-75% by weight) orcombined with partially hydrogenated (i.e., trans-fatty acid-containing)soybean and canola oils. The sunflower oil-rich fats of Moran containvery high levels of linoleic acid (49%-53%) which can undesirablydepress HDL “good” cholesterol. Blair et al. describe margarineoils/fats prepared using about 35%-70% by weight of a palm mid-fractionstructural fat. Their structural fat has a solid fat content (SFC or SFCvalue) that is low, i.e., less than 50% at room temperature (70° F.).Lomneth et al. describe a margarine (spreadable water-in-oil emulsion)in which an oil phase is prepared using a similarly elevated level(35%-70% by weight) of palm mid fraction structural fat with an SFCvalue between 31 and 58% at 70° F. Their structural fat is typicallypartially hydrogenated to decrease its iodine value to 39-50 from itshigher natural IV value of approximately 48 and above. Using palmmid-fraction fats with lower SFC values requires that greater amounts ofthe palm mid-fraction be added to a vegetable oil to achieve hardening.However, these elevated levels of palm mid-fraction undesirably increasethe cholesterolemic effect of the resulting fat blend. McCoy utilizes apalm mid-fraction fat that again has a low solid fat content (from about31% to about 58% SFC at 70° F.). This low SFC necessitates the additionof a large amount of palm mid-fraction, about 35% to 70%, to solidifybetween 30% and 65% of the soft oil. Once again, using such elevatedlevels of palm mid-fraction hard fat contributes high levels ofsaturated fatty acids to a blended fat composition and is expected toraise plasma cholesterol levels when such table spreads are consumed asa nutritional fat on a regular basis.

Thus, there remains a need to develop margarines and table spreads thatdo not raise, and preferably which lower or more effectively lower,plasma cholesterol levels.

SUMMARY OF THE INVENTION

The present invention provides nutritional fat compositions useful forreducing plasma LDL-cholesterol levels and improving the mammalianlipoprotein profile. Compared to fats traditionally consumed in thehuman diet, and hardened fat blends that are high in unsaturated fattyacids, the fat compositions of the invention contain reduced levels ofsaturated fatty acids relative to monounsaturated and polyunsaturatedfatty acids. The compositions of the invention utilize unsaturatedvegetable oils that are blended and hardened with a surprisingly lowlevel of a natural variety of palm mid-fraction (PMF) hardstock fat,which is selected to have a high solid fat content or “SFC” measured atroom temperature (20° C.). This reduced level of PMF contributes aslittle as 9% to 15% by weight of saturated fatty acids (combinedpalmitic+stearic acids) to the total fatty acid content of the fat blendcomposition, thereby beneficially helping to control plasma totalcholesterol levels when the composition is routinely used as anutritional fat. The variety of PMF fat is also selected to contain ahigh level of disaturated triglycerides and a low level of trisaturatedtriglycerides (the latter herein shown to be cholesterolemic), togetherwith the aforesaid elevated solid fat content measured at 20° C. Withfat blend compositions being successfully hardened using a surprisinglysmall amount of PMF, the resulting compositions contain an overallreduced amount of total saturated fatty acids compared to previouslydescribed PMF-hardened fats. The ability to beneficially hardenunsaturated vegetable oils using a low level of saturated fatty acidsand a very low level of trisaturated triglycerides requires utilizing aPMF preparation having both a very high solid fat content (SFC) measuredat 20° C. and negligible SFC (e.g., less than 5% or even less than 3%)measured at 35° C., as well as employing advanced margarine andshortening manufacturing methods. These methods can be combined withphysical, chemical, and mechanical manufacturing conditions that controlcrystallization and hardening conditions, thereby enabling reducedlevels of PMF to form stable solid fat blends with unsaturated naturalvegetable oils such as canola oil, soybean oil and other oils.

Several factors cooperate to provide the LDL cholesterol reducing effectof the hardened vegetable oils described herein. The amount of saturatedfatty acids, which are cholesterolemic, required for hardening anunsaturated vegetable oil has been reduced by decreasing the amount ofPMF hardstock used in the blend. This reduced level of saturated fattyacids is made possible by using a PMF hardstock fat having a high solidfat content due to having a high disaturated triglyceride level (e.g.,high 1-palmitoyl, 2-oleoyl, 3-palmitoyl (POP) triglyceride content) andyet a low trisaturated triglyceride level (e.g., low 1,2,3-palmitoyltriglyceride (PPP, tripalmitin) content. Also important is thecontrolled addition of C18:2 polyunsaturated linoleic acid, from theunsaturated vegetable oil, in the combined fat blend composition. Thatis, a sufficient but not excessive amount of linoleic acid (18:2) isintroduced into the blended fat composition to beneficially reduceplasma total cholesterol levels while not reducing HDL “good”cholesterol levels. A suitable level of linoleic acid ranges between 15%and 45% by weight based on the total fatty acid content of the fatblend. The optimum level of 18:2 within this range depends upon severalvariables, and is preferably determined empirically. The variablesinclude the species and amounts of various saturated fatty acids in thefat blend, as well as the amounts of different saturated fattyacid-containing triglyceride species present in the fat blend.

One aspect of the invention is a method of preparing a blendednutritional fat composition that is hardened with saturated fat. Themethod includes the steps of: (a) selecting at least one palmmid-fraction fat; and (b) mixing the at least one palm mid-fraction fatand at least one unsaturated vegetable oil to form the composition. Thepalm mid-fraction fat is selected to contain from about 60% to about 95%by weight of disaturated triglycerides and less than 6% by weight oftrisaturated triglycerides. In some embodiments, the palm mid-fractionfat contains from about 55% to about 95% by weight of disaturatedtriglycerides. In some embodiments, the palm mid-fraction fat containsless than 5% by weight of trisaturated triglycerides. The triglyceridecomposition of the palm mid-fraction is such that more than 50 mol % (insome embodiments, more than 55%, 60%, 65%, or 70%) of the disaturatedtriglycerides contain either palmitic acid or a combination of palmiticacid and stearic acid at the sn-1 and sn-3 positions and either oleicacid or linoleic acid at the sn-2 position. The fat portion of thenutritional fat composition contains from 10% to 24% by weight of the atleast one palm mid-fraction fat and from 60% to 90% by weight (incertain embodiments from 65% to 90%, or from 70% to 90%, or from 71% to90%, or from 72% to 90%, or from 73% to 90%, or from 73% to 90%, or from74% to 90%, or from 75% to 90%, or from 76% to 90%) of said at least oneunsaturated vegetable oil. The nutritional fat composition also containsfrom 8% to 23% by weight (in some embodiments from about 8% to 20% orfrom about 8% to 18%) of disaturated triglycerides based on the totalweight of triglycerides, and the weight ratio of disaturatedtriglycerides to trisaturated triglycerides in the composition isgreater than 10:1 (in some embodiments greater than 15:1 or greater than20:1). Further, the composition contains from 15% to 45% by weight oflinoleic acid based on the total weight of fatty acids. The fatcomposition may contain intermediate and/or narrower ranges of linoleicacid, e.g., 20% to 40% by weight, 25%-40% by weight, and/or 30%-45% byweight linoleic acid. The sum of lauric acid and myristic acid in thecomposition is less than 10% by weight based on the total weight offatty acids in the composition. In some embodiments, the sum of lauricand myristic acids is less than 9%, or less than 8%, or less than 7% byweight of the total fatty acids. The composition may also containintermediate levels of lauric plus myristic acids, e.g., 1%-5% and/or5%-9% by weight. The composition is solid or semi-solid at 20° C. andfluid at 35° C., and is substantially free of synthetic trans-fattyacids.

Another aspect of the invention is a blended nutritional fatcomposition, which can be, for example, made by the above-describedmethod. The fat portion of the composition contains from 10% to 24% byweight of palm mid-fraction fat and from 60% to 90% by weight (incertain embodiments from 65% to 90%, or from 70% to 90%, or from 71% to90%, or from 72% to 90%, or from 73% to 90%, or from 73% to 90%, or from74% to 90%, or from 75% to 90%, or from 76% to 90%) of unsaturatedvegetable oil. The palm mid-fraction fat contains from about 60% toabout 95% by weight of disaturated triglycerides and less than 6% byweight of trisaturated triglycerides. In some embodiments, the palmmid-fraction fat contains from about 55% to about 95% by weight ofdisaturated triglycerides. In some embodiments, the palm mid-fractionfat contains less than 5% by weight of trisaturated triglycerides. Thetriglyceride composition of the palm mid-fraction is such that more than50 mol % (in some embodiments, more than 55%, 60%, 65%, or 70%) of thedisaturated triglycerides contain either palmitic acid or a combinationof palmitic acid and stearic acid at the sn-1 and sn-3 positions andeither oleic acid or linoleic acid at the sn-2 position. The nutritionalfat composition contains from 8% to 23% by weight (in some embodimentsfrom about 8% to 20% or from 8% to 18%) of disaturated triglyceridesbased on the total weight of triglycerides, and the weight ratio ofdisaturated triglycerides to trisaturated triglycerides in thecomposition is greater than 10:1 (in some embodiments greater than 15:1or greater than 20:1). The composition contains from 15% to 45% byweight of linoleic acid based on the total weight of fatty acids in thecomposition. The fat composition may contain intermediate and/ornarrower ranges of linoleic acid, e.g., 20% to 40% by weight, 25%-40% byweight, and/or 30%-45% by weight linoleic acid. The sum of lauric acidand myristic acid in the composition is less than 10% by weight (or insome embodiments less than 9%, less than 8%, or less than 7%) based onthe total weight of fatty acids in the composition. The composition mayalso contain intermediate levels of lauric plus myristic acids, e.g.,1%-5% and/or 5%-9% by weight. The composition is solid or semi-solid at20° C. and fluid at 35° C., and is substantially free of synthetictrans-fatty acids. In some embodiments, the total content of saturatedfatty acids in the composition is from 15% to 40% by weight, or from 15%to 30% by weight, or from 15% to 23% by weight, based on the totalweight of fatty acids in the composition. In some embodiments, thetrisaturated triglyceride content of the composition is less than 3%, orless than 2%, or less than 1%, based on the total weight oftriglycerides in the composition. In some embodiments, the weight ratioof linoleic acid to total saturated fatty acids in the composition isfrom 1:1 to 3:1. In some embodiments, the solid fat content at 20° C. ofthe composition is from 9% to 24% by weight based on the total weight ofthe composition. Intermediate solid fat content levels at 20° C. thatalso can be used include 9% to 15%, 15% to 20% and 20%-24%. In certainembodiments, the composition contains from 10% to 18% by weight of atleast one palm mid-fraction fat, from 80% to 90% by weight ofunsaturated vegetable oil, and disaturated triglycerides make up from 8%to about 18% by weight of the total triglycerides in the composition. Inother embodiments, the composition contains from 10% to 20% by weight ofat least one palm mid-fraction fat, from 82% to 90% by weight ofunsaturated vegetable oil, and disaturated triglycerides make up from 8%to about 16% by weight of the total triglycerides in the composition. Insome embodiments, the composition consists essentially of palmmid-fraction fat and unsaturated vegetable oil.

Still another aspect of the invention is a prepared food productcontaining a blended nutritional fat composition of the invention. Insome embodiments, the fat composition is blended with water. In someembodiments the fat composition is blended with water to form a foodproduct containing from 15% to 60% water by weight. In some embodiments,the food product contains between 40% and 80% by weight of thenutritional fat composition. In some embodiments, the blendednutritional fat composition is incorporated into a food product selectedfrom the group consisting of margarines, table spreads, shortenings,baked goods, fried goods, filled dairy products, fat-containingconfections, mayonnaise, and salad dressings.

Yet another aspect of the invention is a method of reducing the plasmaLDL cholesterol in a mammalian subject, such as a human. The methodincludes substituting a nutritional fat composition of the invention, ora food product containing such a nutritional fat composition, in theplace of other dietary fat that is solid or semisolid at 20° ′C and isconsumed by the mammalian subject. In some embodiments, the compositionprovides from 10% to 40% of the daily dietary intake of calories by thesubject. In some embodiments, the plasma LDL-cholesterol level isreduced without significantly reducing the plasma HDL-cholesterol levelin the subject.

DEFINITIONS

As used herein, the terms “fat” and “oil” are used interchangeably torefer to an edible triglyceride-based composition. Such fats and oilscan be obtained from a variety of sources, such as plant, microbial, andanimal sources. Fats are generally solid or semi-solid at roomtemperature, while oils are generally fluid at room temperature.

As used herein, “fat portion” refers to the portion of a nutritional fatcomposition that is fats and/or oils and can also include fat- oroil-soluble materials such as monoglycerides, diglycerides, neutralfats, vitamins, or other nutrients. The fat portion does not includewater.

The term “nutritional fat” or “dietary fat” or “triglyceride-baseddietary fat” as used herein means any predominantly triglyceridemolecule-based edible oil or fat, regardless of whether it is derived orpurified from vegetable or animal sources, or is synthetic orsemi-synthetic in origin, or some combination of these. A nutritional ordietary fat may also contain other constituents of choice such asmonoglycerides, diglycerides, flavorings, fat-soluble vitamins,phytosterols and other edible ingredients, food additives, dietarysupplements and the like. While most fats utilized in the compositionsdescribed herein retain their native triglyceride structures (i.e., theyhave not been structurally rearranged or chemically modified), certainof the dietary fat compositions that are discussed herein have beenmodified, e.g., interesterified to rearrange fatty acids (or removecertain fatty acids and attach others) on the glyceryl backbone of thefat. The fat compositions according to the invention can contain smallamounts of chemically modified natural fats or oils, such as thoseproduced by hydrogenation, partial hydrogenation, orinteresterification. Preferably, the compositions do not contain suchmodified natural fats or oils.

The terms “natural fat” and “natural oil” refer to edible fats and oilsthat are extracted from animal, microbial, or plant sources, or apurified fraction of such fats and oils (e.g, a high melting ormid-melting fraction). A natural fat or oil used in the invention issubstantially free of (i.e., possesses less than 10 mol %, less than 5mol %, less than 2 mol %, or less than 1 mol %) of triglyceridemolecules that have been artificially modified in structure (e.g., bychemical or enzymatic means). The edible fat or oil may be from a singlesource or may be a blend from multiple sources.

The term “solid fat content” (SFC) is used in connection with a fat ormixture of fats whose consistency changes from essentially solid toliquid with increasing temperature. In the U.S. the AOCS officialmethods for measuring SFC include AOCS Cd 16b-93 (direct method) andAOCS Cd 16-81 (indirect method). With these methods, the NMR signalsfrom both the liquid and solid components in a fat sample are detectedand measured. More specifically, these methods measure what percentageof all hydrogen nuclei in a test sample composed of hydrogen nuclei inboth liquid and solid phases is due to hydrogen nuclei in the solidphase. The SFC value can be approximated as the percentage by weight ofsolid fat based on the total weight of fat at a particular temperature.That is, the SFC value of a fat composition at a given temperature canbe approximated as the weight of solid fat at that temperature dividedby the total weight of fat (i.e., solid+liquid) in the composition,times 100.

As used in connection with structural changes to fatty acids and/ortriglycerides, the terms “chemically modified”, “synthetic” (e.g.,synthetic trans-fatty acids) and “interesterified” mean that a man-made(i.e., not made by nature) structural change has been introduced thatmodifies the chemical structure of a fatty acid (e.g., by hydrogenation)or the location/site of attachment of a fatty acid in the triglyceridemolecule. Such structural changes may, for example, be accomplished bysynthetic chemical and/or by enzymatic processes.

For some, but not all, of the fat compositions described herein it isbeneficial that the resulting fat-based composition is substantiallycholesterol-free because the presence of cholesterol degrades thelipoprotein profile, undesirably increasing LDL cholesterol andincreasing the LDL/HDL ratio in the plasma. The term “substantiallyfree” in reference to cholesterol level means that the dietary fatcontains less than 10 mg cholesterol per serving of a food containingthe dietary fat, more preferably less than 5 mg per serving, and mostpreferably less than 2 mg per serving to qualify as “cholesterol-free”under current U.S. FDA regulatory standards. Notwithstanding theaforesaid preference, fat compositions described herein are sometimescombined with as much as an equal weight of butter, in which buttercontains 215 mg cholesterol per 100 g, and the resulting fats areencompassed by the present invention. With regard to the definition of“serving size,” the USDA Center for Nutrition Policy and Promotion isthe authority in charge of setting the standards for serving sizes inthe United States. This serving size is referenced on the NutritionFacts label on packaged food products, and allows comparison of thenutritional value of similar as well as different food products. Forexample, milk and yoghurt cite 1 cup or 8 fl. oz., cottage cheese cites½ cup and margarine cites 1 tablespoon as standard serving sizes. TheU.S. Food and Drug Administration (FDA) utilizes the term “ReferenceAmount Customarily Consumed (RACC)” for the amount of a particular foodconsumed by the general population at one eating occasion. If the FDAand USDA serving sizes differ, the USDA serving size is the standardutilized herein.

In reference to fatty acids and their attachment to the glyceryl moietyof the triglyceride molecule, there are three hydroxyl positions foresterification of the fatty acids. These positions allow for differenttriglyceride structural isomers, i.e., stereoisomers to be formed. Thethree points of attachment known as the sn-1, sn-2 and sn-3 positionshave metabolic significance. While the physical properties of the fat(e.g., hardness, melting point crystal structure) are affected by eachfatty acid attached at each position, the fatty acid at the middle orsn-2 position often has the greatest impact on affecting the level ofdifferent plasma lipoproteins. This is because digestion and enzymatichydrolysis by pancreatic lipase removes the sn-1 and sn-3 esterifiedfatty acids, leaving the sn-2 fatty acid monoglyceride to be absorbedinto the bloodstream.

Reference herein to “fatty acids” encompasses both free fatty acids andsuch fatty acids that are esterified to a glycerol backbone in the formof a mono-, di-, or triglyceride. Primarily the fatty acids will bepresent as triglycerides, although appreciable amounts of di- andmonoglycerides may also be present, along with small amounts of freefatty acids.

Indication that a hardstock fat composition such as PMF fat contains a“reduced level or amount of trisaturated triglycerides” means that thefat contains no more than 6% by weight of trisaturated triglycerides onthe basis of the total triglycerides in the PMF fat. In someembodiments, the reduced level of trisaturated triglycerides is lessthan 5% or less than 3% by weight of the triglycerides in the PMF fat.If more than one PMF fat is used, then the above percentage oftrisaturated triglycerides refers to a weight percentage based on thetotal triglycerides in all of the PMF fats taken together. When thehardstock fat is blended and diluted (typically 4-fold or more) with anunsaturated vegetable oil, and the resulting fat composition isidentified as “substantially free of trisaturated triglycerides”, thatmeans that no more than 1.5% by weight of the triglycerides in the fatblend composition contain three saturated fatty acids. In some cases,the percentage will be lower, e.g., no more than 1.25, 1.0, 0.7, 0.5,0.4, 0.3, or 0.2% by number.

Unless indicated to the contrary, range references specified hereininclude the endpoints of the range.

As used herein with respect to percentages or ratios of types of fattyacids in a dietary fat composition, the term “and/or” means that thequantity refers to either one of a pair, or to both of the pairindividually, or to a combination of the pair of specific types orclasses of fatty acids. Each distinct combination of composition levelsis specifically included. For example, specifying that a compositioncontains at least 10% by weight palmitic acid and/or stearic acid meansthat the composition may, for example, contain at least 10% palmiticacid, or at least 10% stearic acid, or at least 10% palmitic acid and atleast 10% stearic acid, or contains a total at least 10% palmitic acidplus stearic acid.

Dietary fat compositions as provided and calculated herein are oftenexpressed in terms of their fatty acid make-up on a weight percentagebasis. As used herein, a percentage of a specific fatty acid by weightrefers to the percentage of that fatty acid with respect to (“based on”)the sum total weight of fatty acids in triglyceride-based fats and fatblends, which is set equal to 100% (not ˜95% as used in certain USDAtables). The ester-linked glyceryl carbon linked to each fatty acid inthe triglyceride molecule is not included in this calculation sincemetabolism and/or hydrolysis of fats yielding fatty acids leaves theglyceryl carbon behind. i

The term “unsaturated fatty acids” as used herein refers to fatty acidscontaining at least one carbon-carbon double bond, and as such, includesall fatty acids except the saturated fatty acids. That is, unsaturatedfatty acids are the sum of monounsaturated and polyunsaturated fattyacids. The most common unsaturated fatty acids include themonounsaturated fatty acid, oleic acid (18:1), and the polyunsaturatedfatty acid, linoleic acid (18:2). The omega-3 polyunsaturated fattyacids include α-linolenic acid (18:3, n-3 or ALA), and the so-calledlong chain omega-3 polyunsaturated fatty acids, eicosapentaenoic acid(EPA) and docosahexaenoic acid (DHA). EPA (20:5, n-3) anddocosahexaenoic acid (22:6, n-3) contain 5 and 6 double bonds in carbonchains of 20 and 22 carbon atoms, respectively.

As used in reference to components in the present compositions, theterms “principally” and “primarily” and “majority” mean that thereferenced component constitutes more than 50% of the indicatedcomposition or combination of components, and in some cases may besignificantly greater, e.g., at least 60, 70, 80, 90 or 95%.

With the consumption of dietary fat as natural triglycerides it isimportant to maintain a ratio of polyunsaturated fatty acids as linoleicacid to saturated fatty acids, in which this ratio can range between 1:1to 3:1 (and sometimes as low as 0.5:1.0), and further in which thedietary fat is substantially cholesterol-free. In reference to thepresent fat compositions, the terms “balanced fat” and “balanced fatcomposition” similarly refer to a fat composition in which the totalsaturated fatty acid content including C12:0 lauric, C14:0 myristic,C16:0 palmitic and C18:0 stearic acid approximates the total C18:2linoleic acid content of the fat composition. When expressed as a ratioof saturated fatty acids to linoleic acid, this ratio can range between0.5:1 and 2:1.

Indication that an edible oil is “rich in palmitate” or “rich inlinoleate” and similar terms means that the oil contains at least 10% byweight, and often at least 15, 20, 25, 30, 40, or 50% by weight of thespecified fatty acids or combination of fatty acids.

As used herein, the term “mammal” refers to both humans and othermammals, and includes experimental mammalian model animals, e.g.,gerbils, hamsters, rats and the like, as well as to livestock animalsreferring to a vertebrate animal which is farmed or ranched. Inparticular, livestock animals include bovines (such as cattle), equines,caprines (such as domestic goats), ovines (such as domestic sheep),avians (such as chickens, turkeys, and ducks), as well as canines andfelines, including companion animals such as domestic dogs and cats.

The term “cholesterolemic effect” as used herein, refers to the abilityof certain foods and food ingredients, including certain fats and fattyacids contained within these fats, as well as cholesterol present inanimal fat, dairy products, meat, fish, and the like, to increase plasmalevels of one or more of the following: total cholesterol (TC), LDL-C,and VLDL-C. Any increase in HDL “good” cholesterol is not included inthis term.

The term “hard fat”, “hardening fat”, “hardstock”, or “solid fat” (inrelationship to triglyceride-based fat compositions) is used herein todescribe a stearin fat, palm mid-fraction (PMF) or palm oil for example,and refers to any edible triglyceride-based fat or fat mixture that issolid at room temperature and that can be combined with one or moreedible liquid oils, e.g., liquid vegetable oils, to solidify these oils,rendering them useful in margarines, table spreads, shortenings andother processed foods. This definition is not meant to exclude the useof other agents that may also be used to solidify edible oils, e.g.,mono- and diglycerides that are closely related to fats.

The term “unsaturated oil”, “unsaturated vegetable oil”, or “unsaturatednatural vegetable oil” as used herein, refers to an edibletriglyceride-based oil or mixture of oils including both monounsaturatedvegetable oils, e.g., olive oil, high oleic sunflower oil and canola oilas well as to polyunsaturated vegetable oils, e.g., soybean oil, cornoil, peanut oil, sunflower oil, safflower oil and cottonseed oil thatare substantially liquid at room temperature.

The term “synthetic trans-fatty acid” or “trans-fatty acid” or“trans-fat” as used herein, refers to a fatty acid, typically within atriglyceride molecule that has been chemically altered by man (usuallyby a partial hydrogenation process) from its natural cis-isomer chemicalbond configuration (between neighboring carbon atoms in the fatty acidstructure) to the so-called trans configuration. Usage of this termherein (carrying the artificial or man-made implication) is intended toexclude naturally occurring trans-fatty acids such as trans-vaccenicacid that is thought to be beneficial and found in the fat of ruminantsand in dairy products, for example.

The terms “trisaturated” (TST), “disaturated” (DST), “monosaturated”(MST) and “unsaturated” (UST) as they modify the term “triglyceride” asused herein, refer to the presence of three, two, one, or zero saturatedfatty acids linked to the 3 carbons forming the glyceride backbone ofthe fat (triglyceride) molecule. These categories of triglyceridemolecules can be denoted as S₃, S₂U, SU₂, and U₃ where S represents asaturated fatty acid, U represents either a monounsaturated or apolyunsaturated fatty acid, and the subscript refers to the number offatty acids (the absence of subscript denoting one fatty acid).

The terms “melting point” (m.p.) and “Mettler Drop Point” as usedherein, are intended to be interchangeable and refer to the transitiontemperature at which a solid fat becomes sufficiently fluid to begin toflow as a liquid droplet out the bottom of a small hole in asample-holding cup. During warming, the Mettler Drop Point is typicallyreached before full melting and optical transparency is achieved. Thesame is true for the softening point (AOCS Method Cc 3-25), the slippoint (Cc 4-25) and the Wiley melting point (Cc 2-38) for fats.

The terms “solid”, “semi-solid”, and “liquid” as used herein, refer tothe physical state of a fat or an oil, mixtures of fats and/or oils,i.e., fat blends, fat-oil blends, oil blends, and any other mixturescontaining fat(s) and/or oil(s) such as margarines, table spreads,shortenings, and other processed foods that include fat(s) and/oroil(s). The term “solid” refers to a material that is firm enough tohold shape under moderate finger pressure. A refrigerated stick ofmargarine or butter is considered solid. By comparison, a table spreadthat is warmed to 20° C., for example, and typically holds its shapeunder the force of gravity, but is very plastic and easily spreadable byknife is considered a “semi-solid.” In contrast, a “solid” material isnot very plastic and is not easily spreadable by knife. A “liquid” or“fluid” material, such as salad oil at room temperature, does not holdits shape under the force of gravity but flows under the force ofgravity.

The following abbreviations are used herein: Solid fat content (SFC);Plasma total cholesterol (TC); Plasma triglycerides (TG); Low densitylipoprotein (LDL); Very low density lipoprotein (VLDL); High densitylipoprotein (HDL); Palm mid-fraction (PMF); Saturated fatty acids (SFA);Monounsaturated fatty acids (MUFA); Polyunsaturated fatty acids (PUFA);Saturated fats (SATS) are fats rich in SFA; Monounsaturated fats (MONOS)are fats rich in MUFA; Polyunsaturated fats (POLYS) are fats rich inPUFA; Palm stearin (PS).

DETAILED DESCRIPTION OF THE INVENTION

The inventors have found that ingesting palm mid-fraction hardstock fatthat contains palmitic and stearic acids at the sn-1 and sn-3triglyceride positions, together with sufficient levels ofpolyunsaturated fatty acids in the form of linoleic acid (C18:2), incombination with other dietary fat, unexpectedly provides beneficialimprovements in the mammalian plasma lipoprotein profile. Withoutintending to limit the invention to any particular mechanism, it isknown that saturated fatty acids increased blood levels of both HDL andLDL, and as a result, ingestion of a diet excessively rich in saturatedfatty acids undesirably elevates total serum cholesterol (TC) and theTC/HDL ratio. However, if a moderate proportion of saturated fatty acidsis ingested together with a sufficient amount of polyunsaturated fattyacids, primarily linoleic acid, the level of LDL is reduced, resultingin a desirably lower TC/HDL ratio.

Surprisingly, it was found that the reduction of TC is more effectivewhen a significant amount of the saturated fatty acids is contributed bya palm oil fraction called palm mid-fraction (PMF) hardstock fat ratherthan other palm oil products such as palm stearin hardstock fat orpartially hydrogenated trans-fatty acid-containing fats, orinteresterified fats containing saturated fatty acids at the sn-2position.

One possible explanation of this effect is that digestion by pancreaticlipase of palm stearin leaves principally sn-2 palmitic acid-containingmonoglycerides, which are cholesterolemic according to the presentinvention, whereas similar digestion of PMF leaves principally sn-2oleic and linoleic acid-containing monoglycerides, which are far lesscholesterolemic according to the present invention. This is illustrated,for example, in the dietary study results shown in the examples below inTables 3 and 6 with the comparison of lipid profiles for a dietcontaining PMF fat (Diet 711) with a diet containing palm stearin (Diet710), or the comparison of a diet containing a PMF-hardened margarine H(Diet 722) with diets containing either a palm kernel oil-hardenedmargarine E (Diet 720) or so-called American Fat Blend margarine B (Diet719). Diets 711 and 722 resulted in notably lower values for TC and/orLDL-C, for example.

When a significant fraction of dietary saturated fatty acids is providedby PMF rather than by palm stearin, and when other cholesterolemiccomponents are not excessive, relatively low levels of linoleic acid canbe effective in limiting TC and LDL levels which might otherwise beelevated from saturated fatty acids contributed by the hardstock fat.This allows relatively low ratios of polyunsaturated fatty acids(primarily linoleic acid) to saturated fatty acids to be effective. Forexample, P/S ratios (in which P is the weight percent of linoleic acidin the fat portion of the composition and S is the sum of the weightpercentages of saturated fatty acids in the same composition) of about1:1 to 3:1 (and also as low as 0.5:1.0) provide sufficient linoleic acidto effectively compensate for the lipoprotein-elevating effects of thesaturated fatty acids.

The animal model data described in the examples below, including plasmalipoprotein data, lead to conclusions that differ from previously heldviews relating to palmitic acid (C16:0) and lipoprotein metabolism.Others have used small increments of milkfat containing sn-2 palmiticacid in dietary fat blends and observed altered lipoprotein profiles inhuman and other mammalian plasmas without suitable explanation. However,in view of the present invention and the data described herein, it isnow apparent that an improved benefit is achieved if two criteria aremet: (1) using PMF hard fat containing predominantly 16:0 and/or 18:0 atthe sn-1 and sn-3 positions to replace less healthy hard fats such aspartially hydrogenated vegetable oils, palm stearin and milkfat (whichupon digestion, yield cholesterolemic sn-2 16:0 and sn-2 18:0); and (2)adding sufficient 18:2 linoleic acid to the blended fat composition byaddition of one or more unsaturated vegetable oils.

A phospholipid derived from an sn-2 saturated fatty acid monoglyceridehas the inherent ability to raise plasma cholesterol levels, especiallyLDL cholesterol (LDL-C). According to the present invention, saturatedfatty acids, especially palmitic acid (16:0), inserted at the sn-2position of triglyceride molecules (present in milkfat as well astrisaturated palm stearin fat), undesirably reduce the HDL-cholesterollevel (HDL-C) and increase the LDL-C/HDL-C ratio. Thus milkfat or palmstearin, in which many or most of the sn-2 positions (approximately 80%in milkfat) are occupied by saturated fatty acid, is the wrong choicefor an advantageous “healthy fat” for adults, principally because itlacks sufficient polyunsaturated fatty acids, such as linoleic acid, atthe sn-2 position. The invention provides dietary fat that is effectiveat improving human lipoprotein profiles and/or glucose metabolism over abroad population by supplying adequate sn-2 unsaturated fatty acids,especially sn-2 linoleic acid.

Furthermore, it is desirable to have no more than a low level oftri-saturated triglycerides in a nutritional fat composition. As aresult, it is beneficial for a large percentage of the triglyceridescontaining one or more saturated fatty acids to also contain at leastone unsaturated fatty acid, preferably including at the sn-2 position asis typically found in natural oils including the PMF hardstock fat ofthe present invention. For example, a beneficial triglyceride may havepalmitate at the sn-1 and sn-3 positions and oleate or linoleate at thesn-2 position. The presence of significant levels of sn-2 unsaturatedfatty acids is further beneficial for the synthesis of phospholipids.These desirable combinations of fatty acids can be provided usingnatural fats and/or oils, without requiring chemicalinteresterification, which generally randomizes fatty acids intriglycerides, resulting in non-natural distributions of fatty acids inthe respective glyceryl positions.

In order to maintain lipoproteins at appropriate levels, certainbeneficial fat compositions contain at most a low or very low level oftri-saturated triglycerides. Such compositions also contain a sufficientlevel of polyunsaturated fatty acids (primarily linoleic acid) to reducethe LDL level. Now, according to the present invention, linoleic acidlocated in the sn-2 position is recognized as most active in thisregard, and therefore it is desirable to provide a sufficient fractionof the linoleic acid located at the sn-2 position. According to theinvention, it is advantageous for only a minor proportion (e.g., lessthan 20 mol %) of the palmitic, lauric and/or myristic acid content in anatural blend of fats and oils to be located at the sn-2 position, butpreferably not at a level that prevents the LDL-lowering effect oflinoleic acid. Preferably, the total amount of lauric and/or myristicacids (i.e., the sum of lauric acid+myristic acid) in the dietary fatcomposition is less than 10% by weight based on the total fatty acids inthe composition. Small amounts of other polyunsaturated fatty acids andsaturated fatty acids may also be present in the composition.Substantially the remainder of the nutritional fat composition ismonounsaturated fatty acids, primarily oleic acid.

Thus, the invention concerns compositions and methods for selectivelyincreasing or at least maintaining the plasma level of HDL “good”cholesterol (HDL-C), decreasing the level of TC and LDL “bad”cholesterol (LDL-C), reducing the TC/HDL ratio, and reducing ormaintaining total blood triglycerides by consuming a substantiallycholesterol-free or limited cholesterol-containing nutritional fat- andoil-based composition that contains an appropriate balance of linoleicacid, oleic acid, and saturated fatty acids, especially includingpalmitic acid. Preferably, the fat composition contains an appropriateamount of unsaturated fatty acids at the sn-2 position, e.g., linoleicacid and oleic acid. The sn-2 unsaturated fatty acids appearparticularly beneficial in this position while also disruptingtri-saturated triglycerides. In some cases, the composition can alsocontain myristic acid and/or lauric acid.

The invention also contemplates fat compositions that assist individualsin limiting total serum triglycerides and/or serum cholesterol andespecially LDL and/or VLDL, and/or limiting the TC/HDL cholesterolratio. These fat compositions can be used in the preparation of foods,as part of prepared foods, and/or directly consumed as part of a diet.

To have a measurable impact on lipoprotein metabolism, the dietary fatblend should be used regularly as a nutritional fat. Many processed foodproducts can be produced, that incorporate the PMF-containing fat blendcompositions taught herein. Among many others, these include baked andfried foods, margarines, table spreads and shortenings that arehardened, i.e., rendered solid or semi-solid at room temperature (i.e.,about 20° C.) with added levels of from approximately 10% to 24% byweight of PMF hardstock fat. Preferably, the hardened nutritional fatcompositions which are solid or semi-solid at 20° C. are liquid at 35°C. and above, so that they melt in the mouth upon consumption.

Nutritional Fat Compositions

A nutritional fat composition according to the invention includes ablend of an unsaturated vegetable oil with a hardstock fat source suchas PMF, which supplies sufficient saturated fatty acids and solid fatcontent to harden the composition at room temperature while maintainingthe cholesterol lowering effect of unsaturated fatty acids provided bythe vegetable oil. PMF invariably contains a certain level oftrisaturated triglyceride molecules (“TST”). These are shown herein tobe cholesterolemic (see below). Consequently, it is important tominimize the total amount of TST added to a blended fat of the inventionvia the addition of PMF by selecting a suitably fractionated PMF. Such aPMF should contain a high level of disaturated triglycerides (“DST”) anda low level of TST. Either a single PMF can be used, or more than onePMF can be combined. For example, PMFs derived from different sourcematerials can be used together, as well as PMFs produced by somewhatdifferent fractionation procedures. Another alternative is to blend oneor more PMFs with selected components, such as desired DSTs, obtainedfrom other (e.g., non-palm) natural sources, made by chemicalmodification of natural fats (e.g., by interesterification), or madesynthetically. The PMF, or combined or supplemented PMF(s), should beblended with at least one unsaturated vegetable oil to provide an amountof polyunsaturated fatty acids, preferably as 18:2 linoleic acid, thatis approximately equal to or greater than the total weight of saturatedfatty acids contained in the blended nutritional fat composition.

The present inventors have previously developed dietary fat blends thatreduce plasma LDL-cholesterol levels in humans and improve theirlipoprotein profiles using palm oil blended with polyunsaturatedvegetable oils (see U.S. Pat. Nos. 5,578,334; 5,843,497; 6,630,192; and7,229,653). These fat blends provide a weight ratio of polyunsaturated(P) linoleic acid (18:2), to total saturated (S) fatty acids ofapproximately 1:1. However, by using PMF as a hardstock fat, thecurrently useful P/S ratio range can be beneficially made considerablybroader, extending upward from approximately 1:1 up to approximately3:1. This is because low levels of certain PMF preparations areunexpectedly effective at solidifying liquid vegetable oils. Instead ofrequiring 25%, 30%, 35% or higher levels (by weight) of PMF included inblended fat compositions, the present inventors found that levels as lowas 15% or 20% by weight PMF can be effective to render a vegetable oilsolid or semi-solid at room temperature. Therefore, unexpectedly greaterand beneficial proportions of polyunsaturated fatty acids can be addedrelative to saturated fatty acids in PMF fat blends.

The question was investigated whether any palm oil fractions arebeneficially less cholesterolemic than others. For example, could any ofthe three glyceride carbon positions (sn-1, sn-2, or sn-3) be apreferred location for carrying the major saturated fatty acids in palmoil, i.e., palmitate (C16:0) and stearate (C18:0), and how does thisaffect lipoprotein metabolism? As a corollary question, with palm oiland its subfractions, if a certain amount of palmitic acid is to beadded to a blended vegetable oil/fat composition to harden the oil,would there be any benefit in providing the palmitic acid as palmstearin versus palm oil versus palm mid-fraction? As shown below, eachof three different palm oil-related fats was found to contain verydifferent proportions of three classes of saturated fattyacid-containing triglycerides, and these were evaluated for theireffects on mammalian lipoprotein metabolism.

In Table A, the percentage content of palmitic acid is provided (“% P”based on a total of 100%) for the three classes of saturated fattyacid-containing triglycerides in palm stearin, PMF, and palm oil. Therelative C16:0 palmitate content is shown in the numerator along withthe relative weight of the saturated triglyceride components (in thedenominator) is shown for each of the fats (weight of trisaturatedtriglyceride (“TST”), disaturated (“DST”) and monosaturated triglyceride(“MST”)). The data provided in the following Table A for selectedsources identified below for palm stearin, palm mid-fraction and palmoil are calculated as follows:

TABLE A % P/weight TST % P/weight DST % P/weight MST Palm Stearin 52/3540/37 8/16 Palm Mid- 5/3 90/84 5/10 Fraction Palm Oil 15/9  61/46 24/35 From these numbers, it is apparent that the major concentration ofpalmitic acid within the classes of saturated fatty acid-containingtriglycerides shifts from principally TST+DST for stearin to almostexclusively DST for palm mid-fraction to a mixture of DST+MST for palmoil.

In the case of a palm stearin of IV 30.5 (IV=iodine value, see Che Manet al., “Composition and Thermal Profile of Crude Palm Oil and ItsProducts,” JAOCS; 76; 237-242; 1999), it is calculated thatapproximately 52% by weight of the palmitate content is found intrisaturated (“TST”), 40% in disaturated (“DST”) and 8% in monosaturated(“MST”) triglyceride molecules, where overall, the stearin containsapproximately 35% by weight TST, 37% DST and 16% MST. By comparison, fora 34.4 IV palm mid-fraction (Moran, U.S. Pat. No. 4,115,598) it iscalculated that approximately 5% by weight of the palmitate content isfound in trisaturated, 90% in disaturated and 5% in monosaturatedtriglyceride molecules, where overall, the mid-fraction containsapproximately 3% by weight TST, 83% DST, 10% MST, and 4% unsaturatedtriglycerides. And finally, for a 51.5 IV RBD palm oil (Che Man et al.,1999), it is calculated that approximately 15% by weight of thepalmitate content is found in trisaturated, 61% in disaturated and 24%in monosaturated triglyceride molecules, where overall, the palm oilcontains approximately 9% by weight TST, 46% DST, 35% MST, and 5%unsaturated triglycerides.

PMF is fractionated from natural palm oil and, among a variety ofsaturated fatty acid-containing triglycerides present therein, PMFcontains predominantly DST molecules that combine saturated fatty acids(palmitate and some stearate) with an unsaturated fatty acid (oleate orlinoleate) typically in the middle (sn-2) position. PMF differssignificantly from palm stearins, which contain an abundance of TSTmolecules in combination with DST molecules. On the other hand, palmolein differs from both PMF and palm stearin by containing largely MSTmolecules. It is difficult to predict a priori which of these fats is“healthiest” in terms of lipoprotein metabolism when blended with anunsaturated vegetable oil such as canola and/or soybean oil. Each solidfat is expected to be metabolized somewhat differently, and when dilutedand co-metabolized with an unsaturated vegetable oil, may yet again bemetabolized differently, to produce varying levels of TC, LDL, VLDLand/or HDL cholesterol in the mammalian plasma.

Different commercial preparations of PMF can vary widely in theirchemical composition and physical specifications, including but notlimited to their iodine values, solid fat contents as a function oftemperature, and their triglyceride structural isomer contents fortrisaturated, disaturated and monosaturated triglyceride molecules. Forthe sake of illustration, it is informative to compare the triglyceridemolecules in two theoretical PMF preparations containing the samepercentages by weight of palmitate and oleate, PMF-1 may contain 3 partsby weight disaturated POP (palmitate at the sn-1 position, oleate atsn-2 position and palmitate at the sn-3 position)+1 part monosaturatedPOO+1 part trisaturated PPP, whereas the second preparation, PMF-2 maycontain exactly the same amounts of various fatty acids, but may contain5 parts by weight of almost exclusively POP. Among the commercialpreparations of special interest for usage herein are PMF preparationsrich in DST, e.g., PMF-2 above. Commercial PMF fats whose solid fatcontent (“SFC”) at 20° C. exceed 50%, 60% or even 70%, but whose SFC at35° C. is low ornegligible are of special interest herein because theytend to be particularly rich in DSTs while being depleted of both MSTand TST (the latter TST having a melting point above 35° C., and shownherein to be undesirable owing to its cholesterolemic nature inmammals).

For the purposes of the present disclosure, important and preferred DSTmolecules in PMF include 1,3-dipalmitoyl-2-oleoyl glycerol (also calledPOP triglyceride), 1,3-dipalmitoyl-2-linoleoyl glycerol (also called PLPtriglyceride) and 1-palmitoyl-2-oleoyl-3-stearoyl glycerol (also calledPOS/SOP triglycerides). Research disclosed herein shows a remarkablebenefit derived from using a minimum but sufficient amount of PMF as ahardening fat in which the PMF selected, contains as small a proportionof trisaturated triglycerides as possible (through appropriatefractionation of PMF from palm oil). Furthermore, the selected PMF fathas a particularly high solid fat content (SFC) at 20° C. owing to thepredominance of DSTs.

Natural palm oil fractions available for hardening a vegetable oilinclude palm stearin, palm mid-fraction and whole palm oil. Among these,there are multiple parameters that may be considered in selecting asolid palm fat. Various palm fractions differ in cost, initial softeningand melting point temperatures, hardening efficacy at a particulartemperature per gram of material (reflecting the SFC at thattemperature), saturated fatty acid content (SFA) and adversecholesterolemic potency per gram. Whole palm oil is cost-effective, butfor many uses has too low a softening temperature to be an effectivehard fat. On the other hand, both palm stearin (m.p. ˜56° C.) andmid-fraction (m.p. ˜32° C.) have been found to be effective hard fatsfor solidifying vegetable oils, and both have found commercial uses.Because the stearin fraction has a higher melting point than themid-fraction, there is an upper limit to the amount of stearin that canbe added to a vegetable oil before the hardened fat acquires a waxymouth feel, whereas more mid-fraction can be added without this problem.With regard to triglyceride structure, palm stearin containsapproximately 35% by weight TST, 37% DST and 16% MST whereas, dependingon fractionation conditions, palm mid-fraction typically contains 5% orless TST, 55%, 60%, 70%, 80% or more of DST, and approximately 10-20%MST. Therefore, the high level of TST present in stearin versus the highlevel of DST present in mid-fraction are important in distinguishingthese palm fractions.

For the sake of comparison, it is interesting to consider adding 10% byweight saturated fatty acids (SFA) from palm stearin or from palmmid-fraction to separate batches of vegetable oil shortening to hardenthe oils. The mid-fraction with its predominant DST population ofmolecules may add up to 50% more saturated fatty acid-rich triglyceridemolecules than the stearin with its predominant TST molecules. Comparingthe likely plasma cholesterolemic response to DST and TST molecules,while it could be argued that a DST molecule is less cholesterolemicthan a TST molecule, the actual lipoprotein results, i.e., TC andLDL-cholesterol levels as well as the ratios of TC (and LDL-C) to HDL-Cin mammalian plasma, are difficult if not impossible to predict withoutexperimentation. Furthermore, the extent to which a particularSFA-containing triglyceride molecule is cholesterolemic may depend uponthe level of other triglycerides in a meal, as well as the levels ofparticular polyunsaturated fatty acid and monounsaturated fatty acidmolecules in the dietary fat. Accordingly, carefully controllednutritional studies have been performed in which all nutritionalvariables were kept substantially constant except for one variable beingtested. In the experiments described below in Example 2 (see Tables 1and 2), the amount of dietary fat (expressed as a % of dietary energy),the ratio of dietary fatty acids (PUFA/MUFA/SFA) and the particularspecies of fatty acids (e.g., palmitic, stearic, etc.) were maintainedrelatively constant, while, for example, the saturated fat-containingtriglyceride structure (DST versus TST) was varied by feedingappropriate levels of palm mid-fraction versus palm stearin.

A preferred fat composition of the invention contains triglycerides withlow saturated fatty acid content at the sn-2 position. The compositioncan contain myristic and/or lauric acid, some of which may be in thesn-2 position, while the amount of sn-2 palmitate should be minimized.Nevertheless, significant amounts of palmitic acid, such as provided byPMF fat, can be safely incorporated into fat compositions provided thatthe palmitate is selectively localized at the sn-1 and sn-3 positions.Conversely, from the results reported herein (see Tables 3 and 6), palmstearin, which contains high levels of tripalmitate and providessubstantial amounts of sn-2 palmitic acid, should be minimized.

The preferred composition also includes sufficient linoleic acid toreduce LDL-C without significantly reducing HDL-C. For example, thecomposition preferably contains linoleic acid at from 15 to 45%, from 10to 40%, from 8 to 40%, or from 9 to 40% by weight based on the totalfatty acid content. The composition also preferably contains saturatedfatty acids at from 15% to 40% by weight based on the total fatty acidcontent, and from 30% to 65% by weight of oleic acid based on the totalfatty acid content. Oleic acid is considered an essentially “neutral”fatty acid in terms of affecting cholesterol, LDL, and HDL. The sum ofweight percentages for saturated, monounsaturated and polyunsaturatedfatty acids in all cases equals 100%. The phrase “without significantlyreducing” HDL-cholesterol means that the HDL-cholesterol level is notreduced by more than 5%. In some embodiments, HDL-cholesterol remainsabout the same, and in some embodiments it may be increased.

The advantage of utilizing a nutritional fat composition according tothe invention as a component or replacement for a significant portion ofa dietary fat is that it can beneficially affect LDL-C and HDL-C levels.There is an extensive body of clinical evidence that dietaryintervention with edible fats and oils that increase plasma levels ofHDL cholesterol, while decreasing the TC, LDL-C, VLDL-C levels, and theratios of LDL-C to HDL-C and TC to HDL-C, all provide substantial healthbenefits in terms of reducing the risk of coronary heart disease andother health problems.

The proportion of polyunsaturated fatty acids, such as linoleic acid, tobe used in a dietary fat composition is also an important consideration.The concept of balanced fatty acids between polyunsaturated andsaturated fatty acids is described by Sundram et al. in a series of U.S.patents cited above, with the proportion of linoleic acid being set from15% to 40% by weight and the total saturates being set from 20% to 40%by weight. According to the present invention, however, the range oflinoleic acid in a dietary fat is from 15 to 45%, from 10 to 40%, from 8to 40%, from 9 to 40% or from 10 to 40% by weight. A preferred range oflinoleic acid is from 10 to 20% by weight, but in some cases as high as30 or 40% can be used, for balancing a total saturated fatty acidcontent of from 15% to 55% (or in particular cases, 15 to 45% or 15 to40%) by weight in the dietary fat composition. The lower levels oflinoleic acid of the present invention are more effective because thelevel of trisaturated triglycerides is also low, and because the levelof sn-2 saturated fatty acids, especially sn-2 palmitate, is low. Thepresent invention teaches that it is not advantageous simply to increasethe level of polyunsaturates, which is surprising and contrary toconventional nutritional teaching, because at excessively high levels,the lipoprotein profile becomes less desirable.

The proportion of saturated fatty acids in the dietary fat is also amatter to be considered in formulating a balanced oil. Clearly, thelevel of myristic acid and/or lauric acid and the proportions ofmyristate and/or laurate residues (including in the bioactive sn-2position of the triglyceride molecule) are important. But it is an openquestion as to the degree to which the overall proportion of saturatedfatty acids in the diet is critical (excluding myristate and/orlaurate). The discussion in U.S. Pat. Appl. No. 20110166227 suggeststhat dietary cholesterol may be a substantial problem when fed withsubstantial levels of saturated fatty acids in degrading a healthylipoprotein profile. Given that saturated animal fats in meats andbutter are widely consumed and widely known to contain substantiallevels of cholesterol, whereas saturated vegetable fats (e.g., palm oil)that are cholesterol-free have been historically less common in theAmerican diet and poorly understood, there is a common misconceptionthat all saturated fat is harmful. In fact, in light of the HDL-C andLDL-C/HDL-C data presented in Table 1 of U.S. Pat. Appl. No.20110166227, it is suggested that the range of total saturated fattyacids in a dietary fat can safely vary between 15% and 40% or even 50%by weight in the substantial absence of dietary cholesterol and in thepresence of adequate unsaturated fatty acids, especially polyunsaturatedfatty acids.

The cholesterol concentration in a dietary fat should be minimized toavoid degrading the lipoprotein profile. It is preferable that dietarycholesterol not exceed 2 mg per serving as this is the maximumpermissible cholesterol level allowed under U.S. FDA regulations for aproduct to be labeled as cholesterol-free. For a 14 g serving of tablespread, this level represents 0.014% by weight cholesterol. Bycomparison, a low cholesterol food may contain up to 20 mg (0.14% byweight) cholesterol per serving, while butter typically contains 0.22%by weight cholesterol (16-fold higher than a cholesterol-free product).In advantageous cases, the fat composition contains no more than 0.014%by weight cholesterol, but in some instances may contain as much as0.12% by weight cholesterol (approximately half the level in butter).

The level of linoleic acid in the nutritional fat composition issufficient for decreasing LDL even when fed in the presence of asubstantial but not excessive level of myristic acid (14:0), lauric acid12:0), or the combination of lauric acid (12:0) and myristic acid(14:0), especially when the level of trisaturated triglycerides is low.Preferably, the levels of palmitic acid (16:0) and stearic acids (18:0)are also low. As little as 15% by weight or less (e.g., about 10-14.9%)of linoleic acid can be sufficient to minimize the LDL/HDL cholesterolratio when the diet contains the appropriate levels of myristic, lauric,or lauric+myristic fatty acids.

Thus, even though the invention includes edible fat compositions whichinclude 15% to 45% linoleic acid, surprisingly advantageous dietary fatcompositions (and food containing such fat compositions) can be preparedwith a PMF hardstock fat such that the fat composition contains lessthan 15% linoleic acid (e.g., 10-12%, 10-14.9, or 12-14.9%). Such fatcompositions also contain from 15 to 45%, 15 to 40, 15 to 30, 20 to 50,20 to 45, 20 to 40, 20 to 35, or 20 to 30% by weight saturated fattyacids. Preferably the total level of saturated fatty acids does notexceed the just stated levels. Also preferably, palmitic acid (16:0)constitutes no more than 25% of total fatty acids by weight, morepreferably no more than 20%, and still more preferably no more than 15,12, or 10% of the total fatty acids by weight. Stearic acid preferablyconstitutes no more than 10%, more preferably no more than 9%, 8, 7, 6,5, 4, or 3% of the fatty acids by weight. Aside from linoleic acid andsaturated fatty acids, substantially the remainder of the fatty acidcontent in the fat composition is preferably oleic acid (18:1) and canalso include minor amounts of other saturated, monounsaturated, and/orpolyunsaturated fatty acids. In certain embodiments, the specified levelof linoleic acid is replaced with a combination of at least 2, 3, or 4polyunsaturated fatty acids selected from the group consisting oflinoleic acid, alpha-linolenic acid, eicosapentaenoic acid (EPA),docosahexaenoic acid (DHA), and any combination thereof, but preferablyat least 3, 4, 5, 6, 7, 8, 9, or 10% by weight is linoleic acid.

Certain desirable fat compositions can be prepared by blending differentfats having appropriate fatty acid profiles. For example, palm kerneloil can be used to provide myristic and lauric acid. Advantageously,many of the triglyceride molecules in palm kernel oil contain sn-2unsaturated fatty acids, but nearly all of these are sn-2 oleic acidrather than the more beneficial 18:2 linoleic acid. Based upon 100% byweight of the fatty acids contained in a fat, palm kernel oil commonlycontains about 49% lauric acid (12:0), about 17% myristic acid (14:0),about 8% palmitic acid (16:0), about 12% oleic acid (18:1), and about2-3% linoleic acid (18:2), along with about 2-4% each of other saturatedfatty acids (stearic 18:0, capric 10:0, and caprylic 8:0). Oleic acidcan be provided, for example, by blending with high oleic sunflower oil,such as that from Cargill Inc. (Minneapolis, Minn.) or the high oleicsoybean oil from DuPont. The Cargill high oleic sunflower oil containsapproximately 82% oleic acid, 8-9% linoleic acid and 8-9% saturatedfatty acids, while the DuPont high oleic soybean oil containsapproximately 84% oleic acid, 3% linoleic acid, and 13% saturated fattyacids. If desired, additional linoleic acid can be contributed by addingany of a variety of vegetable oils containing substantial amounts oflinoleic acid, e.g., standard or commodity soybean, safflower,sunflower, and/or corn oils, or by adding appropriate amounts ofsynthetic triglycerides having linoleic acid at the sn-2 position.

In preferred embodiments and as specified more particularly below, thePMF hardstock composition contains substantial levels of a combinationof palmitic acid and stearic acid, e.g., at least 55% by weight (the sumof fatty acids totaling 100%). In some cases, the composition alsocontains a measurable level of myristic acid or of lauric acid or both.Thus, the composition may contain 0.5% or more by weight of eithermyristic acid or lauric acid or a combination thereof. The compositionfurther contains an appropriate balance of polyunsaturated fatty acids,in particular linoleic acid, usually in a weight ratio ofpolyunsaturated fatty acids (or linoleic acid) to saturated fatty acidsof about 1:1 to 4:1.

Thus, for some embodiments, the ranges for the P/S ratio of 18:2linoleic acid to total saturated fatty acids are provided as follows.Where ranges of numerical ratios are specified, each number is a ratioin which the denominator is 1.00. In the immediately following ranges,the denominator of 1.00 represents total saturated fatty acids. Thus byway of example, a range of 1 to 4 means that the P/S ratio ranges from1:1 to 4:1. Accordingly, the presently described ranges of P/S ratiosare from 0.5 to 4, 0.5 to 3, 0.5 to 2.5, 0.5 to 2, 0.5 to 1.5, 0.5 to 1,0.75 to 4, 0.75 to 3, 0.75 to 2.5, 0.75 to 2, 0.75 to 1.5, 0.75 to 1,0.8 to 3, 0.8 to 2.5, 0.8 to 2, 0.8 to 1.5, 0.8 to 1.2, 0.9 to 3, 0.9 to2.5, 0.9 to 2, 0.9 to 1.5, 0.9 to 1.4, 0.9 to 1.3, or 0.9 to 1.2, 0.9 to1.1, 1 to 4, 1 to 3, 1 to 2.5, 1 to 2, 1 to 1.5, 1.5 to 4, 1.5 to 3, 1.5to 2.5, 1.5 to 2, 2 to 4, 2 to 3, 2.5 to 4, 2.5 to 3; the weight ratioof 18:2 linoleic acid to saturated fatty acids in the edible fat blendcomposition is at least 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.5,1.7, 2.0, 2.5, 3.0, 3.5 or 4.0, or is in a range of 0.5 to 3.0, 0.5 to2.0, 0.5 to 1.0, 1.0 to 3.0, 1.0 to 2.0, 2.0 to 3.0, or 2.0 to 4.0.

Preferably the fat composition includes a palm mid-fraction PMF hard fatin which the weight ratio of oleic acid (18:1) to the sum of palmiticacid (16:0)+stearic acid (18:0) (i.e., the (18:1):(16:0+18:0) ratio,also known as the O/(P+S) ratio) contained in the PMF fat is in a rangefrom 0.45:1 to 0.75:1; the O/(P+S ratio is from 0.45 to 0.70, 0.45 to0.65, 0.45 to 0.60, 0.45 to 0.55, 0.50-0.75, 0.50 to 0.70, 0.50 to 0.65,0.50 to 0.60, and advantageously approximately 0.50:1±0.05. This ratioof oleic acid÷(palmitic+stearic acid) reflects the amounts of variousmolecular species present in different palm oil fractions (see Table14). Thus, a ratio of 0.50:1 is a good indicator that the proportion ofDST molecules (such as POP) is beneficially high.

Preferably the fat composition includes a palm mid-fraction PMF hard fatin which the PMF fat has a solid fat content at 35° C. (SFC35° C.) ofapproximately zero percent, 1%, or at most 2-3% by weight, and an SFC at20° C. (SFC20° C.) of at least 45% by weight but advantageouslyconsiderably higher, such as 80%-90% by weight; the (SFC20° C.) of thePMF ranges from 45% to 95% by weight, 45% to 90%, 45% to 85%, 50% to95%, 50% to 90%, 50% to 85%, 55% to 95%, 55% to 90%, 55% to 85%, 60% to95%, 60% to 90%, 60% to 85%, 65% to 95%, 65% to 90%, 65% to 85%, 70% to95%, 70% to 90%, 70% to 85%, 75% to 95%, 75% to 90%, 75% to 85%, 80% to95%, 80% to 90%, 85% to 95%, or 85% to 90%.

Preferably the fat composition includes a palm mid-fraction PMF hard fatin which the majority, i.e., more than 50%, of the triglyceridemolecules in the PMF hard fat contain either oleic acid or linoleic acidester-linked at the sn-2 position of the glyceride molecule, and containeither palmitic acid or a combination of palmitic and stearic acids atthe sn-1 and sn-3 glyceride positions. This majority of triglyceridemolecules advantageously ranges upward from 50% to at least 55%, 60%,65%, 70%, 75%, 80%, 85% or even as high as approximately 90%.

Preferably the proportions of fatty acids in the overall fat blendcomposition are substantially determined by the choice of unsaturatedvegetable oils selected for the composition, where the PMF hard fatcontributes a maximum of 24% by weight of the total fat (orapproximately 24% by weight of the fatty acids summed to 100%). Sinceadvantageous PMF hard fats contain about 64% SFA per 100% total, themaximum SFA contribution to a blended fat composition is limited toapproximately 0.24×64%=15% SFA. Of this 15% SFA, approximately 90%(i.e., 14%/15%) is palmitic acid. Advantageous PMF hard fats containhigh levels of the POP triglyceride, and such PMFs typically containapproximately 58% palmitic acid and 30% oleic acid, so that with acomposition limit of 24% by weight PMF, the PMF can contribute onlyapproximately 0.24×58% palmitate=14% palmitate and 8% oleate with muchlesser amounts of other fatty acids in the fat composition. Theremainder of the fat composition totaling 76% or more can beadvantageously constituted of unsaturated vegetable oils contributinghigh levels of monounsaturated fatty acids (e.g., canola oil, high oleiccanola oil, high oleic sunflower, high oleic safflower, high oleicsoybean oil) and/or high levels of polyunsaturated fatty acids (e.g.,regular sunflower oil, regular safflower oil, or regular soybean oil) ora combination of both. Given these compositional limits, tests includingtrial and error experimentation has provided useful solid fat blendscontaining beneficially reduced levels of SFA (e.g. less than 20% of the100% fatty acid total) for use in margarines, table spreads andshortenings blends. Furthermore, these blends containing low levels ofSFA can be formulated with approximately equal or up to three- orfour-fold greater amounts by weight of polyunsaturated fatty acids (as18:2 linoleic acid) to offset and balance any cholesterolemic effect ofSFA.

Preferably the PMF hard fat includes approximately 60% by weightpalmitic acid out of a total of 100% fatty acids (in most cases itcontains between 50% and 60% by weight palmitate); preferably the PMFhard fat includes 3% to 7%, 4% to 6% or approximately 5%±1% by weightstearic acid; preferably the PMF hard fat includes 2% to 8%, 3% to 7%,3% to 6%, 3% to 5% or 4%±2% by weight of linoleic acid.

Preferably the PMF hard fat includes approximately 30%-40% by weightoleic acid out of a total of 100% fatty acids; preferably the PMFcontains between 32% and 37% by weight oleate).

In certain embodiments, the combined blended fat composition includesbetween 25% and 65% oleic acid out of a total of 100% fatty acids;preferably the blended fat composition contains approximately 15% to 30%by weight or preferably 20%±5% by weight SFA and between 20% and 50%polyunsaturated fatty acids as linoleic acid, leaving the approximate25%-65% balance for oleic acid as monounsaturated fatty acids. In othercases, a 15%-20% by weight level of SFA may be balanced by an equalamount, or up to a two-fold excess (30%-40%) of polyunsaturated fattyacids as linoleic acid, leaving a balance of either approximately60%-70% oleic acid at the “high end” or a balance of approximately40%-55% oleic acid at the “lower end”. If the level of SFA is increasedto 25% by weight, with between an equal or a two-fold excess ofpolyunsaturated fatty acids (25%-50% linoleic acid), the remainingbalance of oleic acid would range from approximately 25% to 50% byweight.

The fat composition includes at least 5, 10, 15, 20, 25, 30, 35, 40, 50,60, or 70% by weight oleic acid; the fat composition includes 15 to 45%linoleic acid, with, for example, at least 15, 17, 20, 25, 30, 35, 40%or 45% (or in a range defined by taking any two of the just specifiedvalues as end points of the range) or with polyunsaturated fatty acidsin a range as just specified for linoleic acid in which the ratio oflinoleic acid (18:2) to alpha-linolenic acid (18:3) is at least 1.0,1.5, 2.0, 3.0, 4.0, 5.0, 7.0, or 10.0:1.

In certain embodiments, the fat composition includes 15 to 45%, 15 to35, 15 to 30, 15 to 25, 15 to 20%, 20 to 45%, 20 to 40, 20 to 35, 20 to30, 20 to 25%, 25 to 45%, 25 to 40, 25 to 35, 25 to 30, 30 to 45%, 30 to40, 30 to 35%, 35 to 45%, 35 to 40%, or 40 to 45% linoleic acid, or lessthan 45% linoleic acid; and/or the composition includes no more than 45,40, 35, 30, 25, 20 or 15% of palmitic acid or the combination ofpalmitic plus stearic acids, e.g., 15 to 20, 15 to 25, 15 to 30, 15 to35, 15 to 40, 15 to 45, 20 to 25, 20 to 30, 20 to 35, 20 to 40, 20 to45, 25 to 30, 25 to 35, 25 to 40, or 25-45% of palmitic acid or thecombination of palmitic plus stearic acids; the fat composition includesat least 15, 20 25, 30, 35, or 40% by weight palmitic acid or thecombination of palmitic and stearic acids, or contains at least thespecified percentage of each of palmitic acid and stearic acid up to atotal of 15%, 20, 25, 30, 35, 40, or 45% by weight.

In certain cases, in addition to the specified level of 18:2 (n-6)linoleic acid, the fat composition contains one or more otherpolyunsaturated fatty acids taken singly or in any combination from thegroup of omega-3 polyunsaturated fatty acids (providing a combination of2, 3, or 4 polyunsaturated fatty acids) selected from the groupconsisting of alpha-linolenic acid, eicosapentaenoic acid (EPA), anddocosahexaenoic acid (DHA), preferably such combination includes 5-7,5-10, 5-12, 5-14.9, 5-20, 5-30, 5-38, 8-10, 8-12, 8-14.9, 8-20, 8-30,8-38, 10-12, 10-14.9, 10-20, 10-30, 10-38, 12-14.9, 12-20, 12-30, 12-38,15-30, or 15-38% linoleic acid, or other percentage of linoleic acid asspecified above.

In certain embodiments, specifically including those embodimentsspecified above, the fat composition includes no more than 45% saturatedfatty acids (e.g., from 15 to 45% 15 to 40, 15 to 30, 15 to 25, 15 to20, 20 to 45, 20 to 40, 20 to 35, 20 to 30, 20 to 25, 25 to 45, 25 to40, 25 to 35, 25 to 30, 30 to 45, 30 to 40, 30 to 35, 35 to 45, 35 to 40and 40 to 45% by weight saturated fatty acids); palmitic acid (16:0)constitutes no more than 40, 35, 30, 25, 20, 15, 12, or 10% by weight ofthe total fat composition; stearic acid constitutes no more than 10, 9,8, 7, 6, 5, 4, 3, or 2% of the fat by weight; palmitic acid plus stearicacid constitutes no more than 45, 40, 35, 30, 25, 20, 17, 15, 12, 10, 9,8, 7, 6, or 5% of the total fat composition. For the preceding,substantially the remainder of the fatty acids in the fat compositionare preferably oleic acid (18:1) and polyunsaturated fatty acids,usually primarily linoleic acid, and/or in addition to the specifiedlevel of linoleic acid there is present a combination of polyunsaturatedfatty acids as indicated above. Preferably when other polyunsaturatedfatty acids are included, the linoleic acid is at least 15%, 20, 25, 30,35, 40 or 45% by weight of the total fat, e.g., 15%-45% by weight.

In particular embodiments, the edible fat composition includes 15% to45% by weight linoleic acid or other percentage within this range asspecified above, 15% to 40% by weight saturated fatty acids (SFA)consisting principally of palmitic acid and stearic acid, and 30% to 70%by weight oleic acid or a combination of monounsaturated fatty acidswhich are primarily oleic acid, where the total of the fatty acid weightpercentages is 100%.

In particular embodiments, consistent ingestion of the edible fat blendcompositions hardened with a PMF fat as described herein (e.g., as partof a daily diet) may decrease TC or total cholesterol, LDL cholesterol,and/or increases the fraction of total cholesterol which is HDLcholesterol (or decrease TC/HDL), and/or decreases serum triglycerides.For advantageous embodiments, consistent ingestion of the edible fatcomposition results in two or more of the just specified effects takenin any combination.

In certain embodiments, no more than 5, 4, 3, 2, or 1% by weight of thetriglyceride molecules containing saturated fatty acids in the blendedfat composition are tri-saturated triglycerides;

In certain embodiments, the fat composition contains no triglycerideswhich have been subjected to partial hydrogenation or contains no morethan 0.1, 0.2, 0.3, 0.5, 0.7, 1 or 2% by weight triglycerides which havebeen subjected to such hydrogenation.

In certain embodiments, the blended fat composition is a balanced fatcomposition in which the pairwise weight ratios between saturated fattyacids, monounsaturated fatty acids, and polyunsaturated fatty acids isnot greater than 3, 2.5, 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1,1.0, 0.9, 0.8, 0.7, 0.6, 0.5, or 0.4 or is in a range defined by takingany two of the just-specified values as end points of the range; thedietary fat is an essentially unmodified blend of natural fats; thedietary fat also includes about 10 to 24%, 10 to 22%, 10 to 20%, 10 to18%, 10 to 16%, or 15 to 24%, 15 to 22%, 15 to 20%, or 15 to 18% byweight of PMF hard fat having a Mettler drop point higher than roomtemperature, e.g, about 26, 28, 30, 32, 34, 38, 42 or 45 degrees C; andthe PMF hard fat also can include a supplemental saturated fatcontaining a high concentration of disaturated triglycerides, e.g.,cocoa butter or varieties of natural soybean oil containing high stearicacid contents from a high-stearic acid soybean variety, or a highpalmitic acid content fat, e.g., palm oil, or any combination of thejust specified fats or oils, but not stearin fats such as palm stearinor palm kernel stearin that contain high levels of trisaturatedtriglycerides; the blended fat composition has a Mettler drop pointabove room temperature, e.g., above 26, 28, 30, 32, 35, or 38 degrees C.

In particular embodiments, the blended fat composition is a fat blendcontaining or containing about (within ±5, 10, 20, or 25% of the oil andfat percentages) the oil and fat combination described herein in Tables8 and 9; the fat composition is an oil and fat blend containing orcontaining about (within ±5, 10, 20, or 25% of the fatty acidcomposition percentages) the fatty acid composition described herein inTables 8 and 9.

In preferred embodiments, the fat composition is substantially free ofanimal fat or contains no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10% byweight animal fat (e.g., lard, and/or tallow, or a fat fractionthereof), with the exception of butterfat. While it is preferable forthe fat composition to be substantially cholesterol-free, the fatcomposition may contain up 50% by weight butter and therefore contain upto half the cholesterol level found in butter, if butter is thecholesterol source (butter containing approximately 215 mg cholesterolper 100 g). This exception is made in view of the ability of the presentfat compositions to at least partially compensate for the presence ofdietary cholesterol. Therefore, in certain embodiments, the fatcomposition includes some cholesterol, e.g., at least 0.01, 0.02, 0.03,0.04, 0.05, 0.07, 0.10, or 0.12% by weight or is in a range defined bytaking any two of the just-specified values as inclusive endpoints ofthe range.

The fat compositions described above can be employed to limit VLDL, LDL,total cholesterol, and total triglycerides in serum or plasma, even inthe presence of dietary cholesterol. A fat composition of the inventionis considered to “reduce” an LDL-cholesterol level or a totalcholesterol level if any reduction in the level is obtained. In certainembodiments, a reduction in the level of at least 5%, at least 10%, orat least 15% is obtained. Coronary heart disease, and both vascular andgeneral health in humans and other mammals can be improved if the plasmalevels of total cholesterol and LDL are reduced, while the ratio of LDLto HDL is also reduced, and the level of HDL is increased. There is alsoa general consensus, that a low level of plasma triglycerides isbeneficial, and that if the level of very low density lipoprotein (VLDL)also can be reduced, then the triglyceride and lipoprotein levels arebeneficially regulated because HDL is generated, in part, duringcatabolism of VLDL.

The inventors are unaware of any previous nutritional study onlipoprotein metabolism in which PMF and palm stearin were comparedside-by-side with equal percentages of energy provided by fat, and wherean adequate and equal amount of 18:2 was present in both diets tovalidate the comparison between saturated fatty acid exchanges. This isan important point because intake of 18:2 significantly controls all fatmetabolism (Hayes, K. C., Khosla, P. Dietary fatty acid thresholds andcholesterolemia. FASEB J. 6:2600-07, 1992), and it is well establishedthat changing any one of numerous nutritional or physiological variablesin a nutritional fat study can substantially alter the lipoproteinprofile obtained from a group of human subjects or animal models. Thus,changing more than one variable at a time between diets precludesaccurate interpretation of how each variable might contribute to theresponse observed. These variables include, but are not limited to: (a)the age and gender of the animals, (b) the percent of dietary caloriesbeing supplied by fat, as well as by the test fat in question, (c) thepercentages of energy supplied by each saturated fatty acid andpolyunsaturated fatty acids (assuming monounsaturated fat is maintainedconstant), (d) the level of cholesterol supplied in the diet, (e) thedaily total calorie intake, and (f) the sources and relative amounts ofprotein, carbohydrate and fiber in the diet, including the amount ofplant sterols that may be present. The inventors are also not aware ofany nutritional study in which the effects of palm stearin, PMF, andpalm olein on lipoprotein metabolism were compared directly in the samestudy.

Separated fractions of palm oil have been considered essentially“unhealthy” and their associated lipoprotein metabolism has been largelyignored for several reasons. While PMF has been commercially availablefor many decades, it is relatively expensive compared to whole palm oiland is available in more limited supply than palm oil. PMF has beenprincipally utilized in confection manufacture as a less costlysubstitute for cocoa butter. Moreover, palm oil subfractions as well aspalm kernel oil have been considered hypercholesterolemic and to belargely avoided, particularly if simple palm oil (which containspredominantly palmitic acid) is also available as an alternative andless costly saturated fat. Given this history concerning thehyperlipidemic effects of palm fat fractions such as palm stearin, thepresent invention is surprising and unexpected. It would not have beenexpected prior to the invention that PMF, with its high level of solidfat (SFC) at room temperature could function as a healthier dietarysaturated fat than palm stearin. Yet this has been demonstrated by thegerbil studies provided in the examples presented below (see, e.g.,results obtained with Diet 711 compared to Diet 710, and Diet 722compared to Diet 719).

The saturated fatty acids in desirable hardening fats for this inventionare palmitic acid and/or stearic acid, particularly in the form ofsymmetrical disaturated triglycerides with the configurations POP, POS,or SOS, where “P” refers to a palmitate ester, “0” refers to an oleateester, and “S” refers to a stearate ester. Some, most, or substantiallyall of the oleate ester can be replaced with linoleic acid moieties. Insome cases it is desirable to use oil fractions that are enriched in thedisaturated triglycerides and reduced in the trisaturated triglycerides,and often reduced in low melting point triglycerides such astriglycerides which contain one or no saturated fatty acids. In additionto PMF as described herein, some useful hardening fats of these typesare referred to as “cocoa butter equivalents” or “CBEs”, and include,for example, palm oil, cocoa butter, shea butter, sal fat, illipe fat,kokum fat, mango kernel fat, and mid-melting fractions thereof and/orcombinations thereof. Persons familiar with CBEs will recognize avariety of CBEs with a range of physical properties that can be preparedusing different blends of fats and/or oils, or even synthetictriglycerides, having high disaturated triglyceride content. Personsskilled in preparing edible oil fractions will readily understand how tocontrol the compositions of such fractions by controlling thefractionation conditions (e.g., melting or freezing separationtemperatures used) to provide a desirable oil fraction high indisaturated triglycerides and low in trisaturated triglycerides.

To have a measurable impact on lipoprotein metabolism, the dietary fatblend should be used periodically or regularly as a nutritional fat.That is, the dietary fat blend can be used in the diet as a nutritionalfat from time to time, and preferably on a regular basis, such as atleast daily, or more than once daily, or at least several times perweek, such as 2, 3, 4, 5, or 6 times per week. The schedule ofconsumption of the nutritional fat on a regular basis may vary somewhataccording to the needs or desires of the subject.

Many processed food products can incorporate the PMF-containing fatblend compositions taught herein. These include baked and fried foods,margarines, table spreads and shortenings that are hardened, i.e.,rendered solid or semi-solid at room temperature (i.e., about 20° C.)with added levels of from approximately 10% to 24% by weight of PMFhardstock fat, based on the food item's total fat content equaling 100%.PMF typically contains a certain level of trisaturated triglyceridemolecules. These are shown herein below to be cholesterolemic.Consequently, it is important to minimize the total amount oftrisaturated triglycerides added to a blended fat via the addition ofPMF by selecting a suitably fractionated PMF. Such a PMF should containa maximum level of disaturated triglycerides and a minimum level oftrisaturated triglycerides. The PMF should be combined and supplementedin a blended fat composition with at least one unsaturated vegetable oilto provide an amount of polyunsaturated fatty acids as 18:2 linoleicacid that is approximately equal to, or greater than the total weight ofsaturated fatty acids contained in the blended composition. Previouslydeveloped dietary fat blends can reduce plasma LDL-cholesterol levels inhumans and improve their lipoprotein profiles using palm oil blendedwith polyunsaturated vegetable oils (see U.S. Pat. Nos. 5,578,334,5,843,497, 6,630,192, and 7,229,653). Those fat blends use a weightratio of polyunsaturated (P) linoleic acid (18:2), to total saturated(S) fatty acids of approximately 1:1. However, by using PMF as ahardstock fat according to the present invention, the useful P/S ratiorange is considerably broader and higher, extending from approximately1:1 to approximately 3:1. This is because the present inventionindicates that low levels of certain PMF preparations are unexpectedlyeffective at solidifying liquid vegetable oils. Instead of requiring25%, 30%, 35% and higher levels (by weight) of PMF included in blendedfat compositions, levels as low as 15% and 20% by weight PMF can beeffective. Therefore, greater proportions of polyunsaturated fatty acidscan be added relative to saturated fatty acids in the PMF fat blends ofthe present invention.

In certain embodiments, the prepared food product is a cooking oil/fat,an oil spread (e.g., a margarine or table spread), a shortening, a saladdressing; a barbecue or dipping sauce or other condiment, a baked good(e.g., bread, tortilla, pastry, cake, cookie, or pie), or a dairyproduct (e.g., a milk, yoghurt, or cheese); in certain embodiments, thepresent edible fat composition is 2 to 10, 5 to 15, 10 to 30, 30 to 50,or 50 to 100% by weight of the prepared food product.

Another related aspect concerns a human diet or diet formulation whichis intended for, or which when regularly ingested has the effect ofdecreasing the LDL cholesterol, increasing the fraction of HDL versustotal cholesterol, reducing the triglyceride level, and/or decreasingthe LDL/HDL concentration ratio in human plasma, and/or decreasing thefasting blood glucose concentration, in which a substantial amount,e.g., 10 to 100%, 10 to 90%, 10 to 80%, 10 and 75%, 10 to 50%, 20 to100%, 20 to 80%, 20 to 60%, 30 to 100%, 30 to 80%, 50 to 100%, or 50 to80% by weight of the daily dietary fat is provided by the edible fatcomposition of the first aspect, or an embodiment thereof, or isotherwise described herein for the present invention.

In particular embodiments, a human diet formulation is provided inpackaged liquid form or in other packaged form (for example, packed insingle meal or daily meal plan form), e.g., indicated for nutritionalsupplementation or replacement, such as for elderly patients or patientswith compromised digestive systems, and/or for improvement of apatient's lipoprotein profile.

An aspect of the invention is a method of aiding a person to decreasetotal cholesterol levels, increase the percentage of HDL cholesterol asa fraction of total cholesterol, decrease the LDL cholesterol, decreasetriglyceride level, and/or decrease the LDL/HDL cholesterol ratio intheir plasma, and/or decrease the fasting blood glucose concentration.The method involves providing a dietary fat composition according to thefirst aspect above or otherwise described herein for the presentinvention. Preferably the dietary fat composition is substantiallycholesterol-free or alternatively low in cholesterol.

Preferably the dietary fat composition is a blend of natural fats andoils and does not contain trans-fatty acids.

In particular embodiments, the edible fat blend composition is asdescribed for the first aspect above or an embodiment thereof orotherwise described herein for the present invention; the edible fatblend composition is provided at least in part or primarily in one ormore prepared foods or diets or diet formulations (e.g., liquid dietformulations) as specified for an aspect above or an embodiment thereof.

In certain embodiments, the person or subject consuming a nutritionalfat composition of the invention suffers from high TC, high LDLcholesterol and/or from a high TC/HDL cholesterol ratio, and/or a lowpercentage of HDL cholesterol as a percentage of total cholesterol intheir plasma.

Similarly, another related aspect concerns a method of increasing theconcentration or percentage of HDL cholesterol relative to totalcholesterol, decreasing the LDL cholesterol, and/or decreasing theTC/HDL or LDL/HDL cholesterol ratio, decreasing triglycerides, and/ordecreasing the fasting blood glucose concentration, in the plasma of ahuman subject. The method involves consistently ingesting a dietary fatcomposition of the first aspect above or an embodiment thereof or isotherwise as described herein for the present invention.

A further aspect concerns a method of preparing an edible fat blendcomposition by blending at least one PMF hardstock fat with at least oneother edible oil (e.g, 2, 3, or 4 different oils) in proportions suchthat a blended edible fat as described for the first aspect above or anembodiment thereof is formed. Preferably the blended edible fat issubstantially cholesterol-free.

In certain embodiments, the blended edible oil formed is semi-solid at25, 27, 30, 32, or 35 degrees C.

In some embodiments, the fat composition is formed by blending a higholeic vegetable oil (such as canola oil, high oleic canola oil, higholeic sunflower oil, high oleic safflower oil or high oleic soybean oil)with at least one PMF fat.

A further aspect is a method for limiting plasma triglycerides (TG), LDLcholesterol, and/or VLDL cholesterol, and involves providing foringestion a dietary fat that is solid or semi-solid at 20° C., thatincludes between 10% and 24% by weight PMF hardstock fat.

In some embodiments, the dietary fat provides from 10 to 50%, 10 to 40%,10 to 30%, 20 to 50%, 20 to 40%, 20 to 35%, 20 to 30%, 25 to 50%, 25 to40%, 30 to 50%, 30 to 40%, or 35 to 45% of the total dietary calories inthe diet; regular ingestion of the dietary fat reduces plasma totalcholesterol (TC) without significantly reducing HDL; regular ingestionof the dietary fat reduces plasma VLDL and/or LDL cholesterol withoutsignificantly reducing HDL; regular ingestion of the dietary fat reducesplasma triglycerides (TG) without significantly reducing HDL; regularingestion of the dietary fat significantly reduces VLDL and TC withoutsignificantly reducing HDL; regular ingestion of the dietary fat reducesthe LDL/HDL ratio; and/or the specified reductions occur in the presenceof dietary cholesterol.

In desirable cases, the dietary fat is a balanced fat composition inwhich the pairwise weight ratios between saturated fatty acids andpolyunsaturated fatty acids is not greater than 2, 1.9, 1.8, 1.7, 1.6,1.5, 1.4, 1.3; the dietary fat is an essentially unmodified blend ofnatural fats; the dietary fat also includes about 10 to 24%, 10 to 20%,10 to 15%, 15-22%, 15 to 20%, 15 to 18% by weight of a PMF hard fathaving a Mettler drop point of about 30, 32, 35 or 40 degrees C.; and/orthe hard fat includes 10%, 20%, 30%, 40% or 50% palm oil and/or ahigh-stearic acid content fat such as cocoa butter that does not containsubstantial levels of trisaturated triglycerides; the dietary fatcomposition has a Mettler drop point above 26 degrees C., e.g., 28, 30,32, 35, 37, or 40 degrees C.

In some embodiments, the dietary fat is in the form of a margarine, aspread, a shortening, or a frying oil composition; the dietary fat isincluded in a prepared food, e.g, baked goods, filled milk, mayonnaise,salad dressing, or filled yoghurt, or is included in a complete dietcomposition.

A related aspect concerns a method of limiting (which may be reducing)the concentration of TC, LDL and/or VLDL cholesterol in the plasma of ahuman subject consuming a diet containing cholesterol, by consistentlyingesting a dietary fat as specified for one of the preceding aspects,where the dietary fat provides 10 to 50 percent of the total dietarycalories of the subject (or other percentage of the dietary calories asspecified for an aspect above).

In particular embodiments, the dietary fat is as specified for anembodiment of the preceding aspect; the dietary fat is provide in one ormore prepared foods; the dietary fat is provided in a complete dietcomposition; the dietary fat provides 20 to 50%, 30 to 50%, 20 to 40%,25 to 40%, or 35 to 45% of the dietary calories of the subject; thedietary fat is a blend of at least two, three, or four fats and/or oils.

Also in particular embodiments, the animal or subject ingesting thenutritional fat composition is a mammal, a livestock animal such aporcine, a bovine (typically cattle), an ovine (such as domestic sheep),a caprine (such as domestic goat), an equine, a canine (such as adomestic dog), a feline (such as a domestic cat), or a human.

In a related aspect, the invention further concerns a blended fatcomposition (i.e., a blend of at least two fats and/or oils) asdescribed in the first aspect or otherwise described herein for thisinvention.

In certain embodiments, the dietary fat of the present inventionprovides 10 to 50%, 10 to 40%, 10 to 30%, 20 to 50%, 20 to 40%, 20 to30%, 25 to 50%, 25 to 40%, 30 to 50%, or 35 to 45% of the dietarycalories in the diet.

Likewise, the invention concerns a prepared food product which includesa dietary fat as specified in any of the preceding aspects; the preparedfood product may contain cholesterol, e.g., at level as specified in anaspect above.

In certain embodiments, the prepared food product is a baked good, afilled milk, a mayonnaise, a salad dressing, or a filled yoghurt.

Yet another aspect concerns a method of preparing a dietary fat,involving blending an edible fat or oil with at least one PMF hardstockfat, thereby forming a blended dietary fat composition as described foran aspect above or otherwise described herein for the present invention.

In certain embodiments, the dietary fat is as specified for anembodiment of any of the preceding aspects.

In particular embodiments, saturated fatty acids in the blended dietaryfat are largely provided by PMF fat or about 20 to 40, 25 to 45, 30 to50, 35 to 55, 40 to 60, 50 to 70, or 60 to 80% by weight of thesaturated fatty acids in the fatty acid composition are provided by PMFfat; the blended dietary fat is a chemically unmodified blend of naturalfats and/or oils.

Palmitic and Stearic Acids Provided by DSTs from PMF

For achieving the cholesterol-lowering benefits as described herein,between approximately 5% and 24% by weight, and more typically 10%-22%by weight, or 10%-20% by weight, or 5%-15% by weight, or 8%-18% byweight of a suitably selected PMF fat preparation rich in DST, i.e.,containing between 50% and 95% DST, and suitably limited amounts of bothTST and MST (less than 5% by weight TST and preferably less than 4% or3% TST, and less than 25% by weight MST and preferably less than 20%,15% or 10% MST) should be combined with an unsaturated vegetable oil(such as canola oil, soybean, sunflower, safflower corn, peanut, orcottonseed oil) and used as a principal source of dietary fat. Mammalianfeeding experiments that utilize the gerbil animal model are describedherein below. These tests have proven highly predictive of the humanlipoprotein response to dietary fats. When PMF-hardened vegetable oilswere provided as the principal source of dietary fat over the course of4 weeks, plasma LDL-cholesterol levels were surprisingly diminishedcompared to animals fed essentially the same amount of the same fattyacids (palmitate and stearate) provided by palm stearin rather than PMF.PMF fats were utilized in which most TST had been removed byfractionation to enhance the mammalian plasma lipoprotein response toDSTs, e.g., POP, PLP and POS molecules. Commercial palm mid-fractionsare so-named because they crystallize (or re-melt) at an intermediate or“mid-temperature” between the lower melting olein fractions (rich inmonosaturated triglycerides or MSTs) and the higher melting stearinfractions (rich in TSTs). Evidence provided below indicates that byreplacing an amount of palmitic and stearic acids provided in a palmstearin fraction with an approximately equal amount of these fatty acidsprovided in PMF fat, the lipoprotein profile is improved.

Use of PMF in Confections and Margarine

PMF has found uses in the confectionary industry, in which costly cocoabutter has been replaced with less costly PMF that has similar meltingcharacteristics and mouth feel. However, PMF is still 2-3 times morecostly than palm oil so that its use outside confectionary applicationshas been limited. Unless there is a strong reason for using PMF, a morecost-effective palm oil or a suitable amount of trisaturated palmstearin would be used in its place. With regard to margarine use (seeBackground above), Moran and other investigators describe water-in-oilemulsions containing PMF. For example Moran in U.S. Pat. No. 4,115,598describes table spreads containing 60% of an aqueous phase and 40% of afat phase, in which the fat phase contains higher levels, i.e., 25%-30%by weight, of PMF fat that is combined with 70%-75% sunflower oil. Suchwater-in-oil emulsions as confections and margarines can be formulatedas nutritional fat compositions according to the present invention.

Design of Blended Fat Compositions

Natural palm oil stearin (PS) fractionated from palm oil contains asubstantial proportion of trisaturated triglycerides (tripalmitin),i.e., PPP, which has an elevated melting point and, depending upon thelevel used, now appears to be a cholesterolemic fat based upon thegerbil data in Table 3 below (cf. TC and LDL-C for Diet 710 versus Diet711). These factors limit the amount of palm stearin that can be addedto vegetable oil without the resulting mixture acquiring a waxy mouthfeel and/or being deemed unhealthy.

Thus, if a solid fat content (abbreviated “SFC”) of approximately 10% ormore measured at room temperature is required to produce a firm tablespread, and a level of PS in excess of 5% produces a waxy mouth feel,then the level of PS that may be useful is limited. On the other hand,advantageously blended fat compositions can be formulated containingeither low or moderate levels (e.g., 5%-15% or even 20%-24% by weight)of a palm mid-fraction solid fat (abbreviated PMF) that is rich indisaturated triglycerides (DSTs). It is also possible to design blendedfat compositions in which limited amounts of PS (e.g., 3%-5%) arecombined with moderate levels of PMF (e.g., 10%-15%). PMF can befractionated in substantial quantities from natural palm oil. It is richin the POP triglyceride structure in which the sn-1 and sn-3 isomericpositions are principally occupied by palmitic acid and the sn-2position is occupied by oleic acid, and to a lesser degree linoleicacid. Approximately 46% of palm oil triglycerides are DSTs including 30%POP, 9% PLP and 7% POS (P=palmitate, O=oleate, L=linoleate andS=stearate). Interestingly, with cocoa butter in which the stearic acidcontent (33%) is much higher than in palm oil (4%), the DST populationshifts to predominantly SOS and POS where one or both palmitic acids inPOP are replaced by stearic acid. As for selecting a commercial PMFmaterial with a useful SFC level, that level should be greater than 50%at 20° C., and is preferably as high as 75%, 85%, or even higher at 20°C., yet is essentially 0% at 35° C. The latter number is importanttogether with the melting point for mid-fraction palm oil DSTs ofapproximately 32° C. because they assure that a table spread orshortening fat that is solid at room temperature will also melt in one'smouth and not produce a waxy mouth feel. One example of a DST-rich palmmid-fraction hard fat (PMF) is Palmel 35 produced by Fuji Vegetable OilInc. (Savannah, Ga.). It has a remarkably elevated SFC value of 86% at20° C. and a melting point (Mettler Drop Point) of 32° C.

Hardening Efficiencies for Different Solid Fats

Since a major objective of the present invention is to form natural fatblends that are solid at room temperature using a minimum but sufficientamount of saturated fatty acids, an “efficiency factor” at 20° C.(abbreviated “EF”) is herein defined for a saturated fat that is solidat 20° C. The EF value for the fat is calculated from the ratio of thefat's solid fat content at 20° C. (SFC) to the fat's percentage ofsaturated fatty acids (% SFA), and provides a good indication of therelative ability of saturated fatty acids in a particular fat to hardena vegetable oil at 20° C. In the interest of limiting the level of SFAin fat blends and thereby limiting plasma TC, VLDL and LDL cholesterollevels, solid fats that possess increased EF values have been tested. Inthis regard, palm stearin (PS) contains principally trisaturatedtriglycerides, and is an interesting example. A palm stearin productfractionated by Fuji Vegetable Oil, Inc. and having an iodine value of30 has a melting point (Mettler Drop Point) of 56° C. With a SFC ofapproximately 70% for the PS at 20° C. and SFA content of 71%, the EF isapproximately 1. By comparison, a PMF product (also from Fuji) that isrich in disaturated triglycerides is known as Palmel 35 (iodine value of34), has a SFC at 20° C. of 86% with a SFA of 64%. Therefore, theresulting EF is 86%/64%=1.34. Again, by comparison, the EF for regularwhole palm oil (SFC=20-25% at 20° C. and SFA content=51% by weight) isonly 0.40-0.50. This considerably lower EF value for palm oil comparedto PMF (cf. 0.5 versus 1.34) would require the use of much largeramounts of palm oil and higher concentrations of cholesterolemic SFA tosolidify a liquid vegetable oil when compared to PMF and/or PS.

Disaturated Versus Trisaturated Solid Palm Fat Fractions as HardstockFats

Over recent years, a combination of monosaturated, disaturated andtrisaturated triglycerides provided in palm oil have proven useful forhardening liquid vegetable oils. There are specialized food applicationsthat require the use of one palm oil fraction or another, e.g., forreasons of physical stability, chemical stability, melting point ortexture. However, when more than one palm oil fraction is suitable foruse in a food product such as a shortening, margarine, or table spread,Applicant hypothesized that one palm fraction might prove healthier,i.e., less cholesterolemic, than another for the usage levels deemedfunctional and appropriate for a particular product application.Accordingly, given the different EF values for PS and PMF (approximately1.0 versus 1.3 respectively), it would be instructive to compare themammalian lipoprotein responses to each of these different solid fatfractions and determine whether one fat is less cholesterolemic thananother. To make the comparison valid, approximately equal amounts ofSFA from each of these palm fat fractions should be fed to test animals(e.g., gerbils) along with equal amounts of PUFA to maintain the P/Sratio in the diet approximately constant.

The rationale for comparing the lipoprotein response to dietary intakeof PS and PMF is as follows: PS contains largely trisaturated anddisaturated triglycerides while PMF contains primarily disaturatedtriglycerides. Therefore, if the dietary level of SFA is kept constant,the number of trisaturated PS molecules being ingested will be onlyapproximately ⅔ as great as the number of disaturated PMF molecules. Onthe other hand, disaturated PMF triglyceride molecules contain mainly amonounsaturated fatty acid, oleic acid, in the sn-2 molecular position.This fatty acid would persist in the monoglyceride molecule afterdigestive lipase enzymes have cleaved fatty acids at the sn-1 and sn-3positions, and might be less cholesterolemic than the correspondingmonoglycerides from PS digestion. While alternative hypotheses may beoffered, a mammalian animal model feeding experiment with lipoproteinanalysis or a human clinical study is the only meaningful method fordetermining whether PS or PMF is a healthier hard fat when it iscombined with one or more suitable liquid vegetable oils. The gerbil hasproven to be an excellent model over many years of testing forpredicting human lipoprotein responses to fats.

Palm stearin fractions from palm oil contain approximately 71% by weightsaturated fatty acids with a SFC value at 20° C. of approximately 70%.Therefore, the EP is 70%/71%=0.99. This number is surprisingly less thanthe 1.34 value for PMF Palmel 35 described above. With an elevatedmelting point ranging between approximately 54° C. to 62° C. compared to31° C. to 33° C. for the PMF, the palm stearin may contributeundesirable waxy mouth feel if more than 5% by weight of the stearin isadded to a vegetable oil. While interesterified palm oil (IE palm oil)prepared, for example, using palm olein and exemplified by SansTrans™ HF(Loders Croklaan, Channahon, Ill.) has a useful intermediate meltingpoint of 38-44° C., the SFC at 20° C. for this IE vegetable oil is onlyapproximately 30%. While this is substantially higher than that of bothnative palm olein and palm oil, lipoprotein metabolism may beundesirably altered by synthetic rearrangement of fatty acids in thetriglyceride molecule. This alteration is suggested in Table 3 where TCand LDL cholesterol levels for IE palm olein are compared with those fornative palm olein. While statistical significance was not achieved, allof the lipoprotein indices trend downward for the IE palm olein comparedto the native product. With regard to further advantages accompanyingthe use of PMF hard fat, Applicant shows (Table 3) that PMF as ahardstock fat is less cholesterolemic on a gram for gram basis than palmstearin used in hardening many vegetable oils for producing shorteningsand table spreads. This discovery was made using the gerbil mammalianmodel system that has proven to be a reliable indicator of humanlipoprotein metabolism. Also, PMF exhibits little or no waxy mouth feelowing to its melting point (approximately 32° C.) that is below thetemperature of the human mouth.

In still further advantageous compositions, a beneficial balance betweenthe sum of palmitic acid plus stearic acid (representing most of thetotal saturated fatty acids in palm oil and PMF) and the amount ofunsaturated fatty acids as 18:2 linoleic acid in the fat composition areprovided. Notably, it was discovered that the effective level oflinoleic acid in an edible fat composition can be quite low (e.g., 5, 6,7, or 8% by weight or more) when the oil contains sufficiently lowlevels of cholesterolemic components (e.g., trisaturated triglycerides)which, if elevated, could promote formation of LDL cholesterol orotherwise undesirably impact the relative levels of the variouslipoprotein classes and/or total triglycerides. However, the effectivelevel of linoleic acid required for balancing saturated fatty acids ishigher when the level of trisaturated triglycerides and/or otherLDL-promoting components are higher. These compositions are furtheradvantageous in compensating for dietary cholesterol.

EXAMPLES Example 1 Dietary Fat Testing Protocol

The following procedure was used as a model system to evaluate theeffects of different dietary fat compositions. The procedure is similarto that described in Hayes et al., 2004, J Nutr 134:1395-1399). Themodel can be used to determine the effects of nutritional fat or oilcompositions on plasma lipid profile, adipose deposition, and bloodglucose levels. The gerbil model has been confirmed to mimic human plamalipoprotein and total cholesterol responses to dietary fat andcholesterol. See Pronczuk et al., FASEB J. 8, 1191-1200 (1994).

Gerbils were obtained from Charles River Labs (Wilmington, Mass.) at50-55 g body weight. Animals were housed individually in a laboratoryfacility with conditioned air maintained between 68-72 degrees F. with12 h light/dark cycle. Gerbils were fed a purified diet as described byHayes et al., where total nutrients including carbohydrate, fat, andprotein are standardized and held constant, with appropriate vitamin andmineral mixes. Only the fat composition was manipulated, while fatitself was fed at approximately 40% of the calories, as described in thetables below. Food was fed ad libitum and weighed daily, while thegerbils were given free access to water. Body weights were measuredweekly for 4 weeks, at which time animals were exsanguinated underanesthesia.

The plasma was separated by conventional methods, and lipoproteins wereisolated by density-gradient ultracentrifugation as VLDL, LDL, and HDLusing the method described in Chapman et al., 1981, J Lipid Res22:339-358. Total plasma and isolated lipoprotein fractions were assayedfor cholesterol with a standard enzymatic kit. Plasma triglycerides weresimilarly measured with a conventional enzymatic kit configured for thatpurpose.

Appropriate control diets were included, e.g., diets providing extremesin levels of saturated fatty acids versus linoleic acid, therebyproviding standards for determining the effects on lipid/lipoproteinlevels in response to test dietary fat compositions. For example, in onecontrol diet the dietary fat was provided by an oil high in saturatedfatty acids and low in linoleic acid, and in a second control diet thedietary fat was provided by an oil low in saturated fatty acids but richin linoleic acid (18:2) (safflower oil). Additional intermediate controldiets were also utilized, such as a diet in which the dietary fat wasprovided by a high oleic acid oil with approximately equal levels oflinoleic acid and saturated fatty acids (olive oil).

Example 2 Gerbil Model Lipoprotein Studies with Palm Oil Fractions

The gerbil model described in Example 1 was used to test differentfractions of palm fats. The gerbil model has previously proven to behighly predictive of human plasma lipoprotein responses to dietary fats.

With each of four groups of gerbils (see Tables 1 and 2), elevateddietary levels of saturated fatty acids (SFA) (palmitic acid was 40-60%of total fatty acids were provided either by palm stearin (diet 710), bya PMF preparation (diet 711), by palm olein (diet 713), or byinteresterified palm olein (diet 714). For all groups of animals, 41% ofdietary energy was provided by fat, with SFA accounting for 27%, 24%,19%, and 19% of energy respectively (Table 2). Polyunsaturated fattyacid content was maintained constant at a relatively low level of 4.6%of energy to allow the relative effects of the different saturated fatsto be maximally expressed.

TABLE 1 Gerbil diet compositions with palm oil fractions Diet: 710-714Diet 710 711 713 714 Palm Fraction Pst PMF-35 Pol IE Pol CHO:Fat:Protein(% E) 41:41:18 41:41:18 41:41:18 41:41:18 Kcal/g 4.3 4.3 4.3 4.3 DietaryIngredient (g/kg) Casein 100 100 100 100 Lactalbumin 100 100 100 100Dextrose 200 200 200 200 Cornstarch 174 + 60 (w/gel) 174 + 60 (w/gel)174 + 60 (w/gel) 174 + 60 (w/gel) Cellulose 100 100 100 100 Fat:(SFA:MUFA:PUFA as % E) (27:9:4.6) (24:12:4.6) (19:17:4.6) (19:17:4.6)(P/S) (0.17) (0.19) (0.25) (0.25) Palm Stearine (Pst) 185 0 0 0Palmel-35 (PMF-35) 0 180 0 0 Palm Oil (PO) 0 0 0 0 Palm Olein (Pol) 0 0200 0 IE Palm Olein (IE Pol) 0 0 0 200 Safflower Oil 15 20 0 0 Mineralmix 50 50 50 50 (Ausman - Hayes) Vitamin mix 12 12 12 12 (Hayes -Cathcart) Choline chloride 3 3 3 3 Cholesterol 0.8 0.8 0.8 0.8 Combine60 g cornstarch with 800 mL water to produce gel that is added to theremaining dry ingredients

TABLE 2 Fatty acid profile of dietary fat Diet 710 711 713 714 PstPMF-35 Pol IE Pol Fatty acid % 8:0 + 10:0 0.0 0.0 0.0 0.0 12:0 0.2 0.10.3 0.3 14:0 1.1 0.6 1.0 1.0 16:0 60.2 52.4 40 40 18:0 4.8 5.3 4.3 4.318:1 22.2 30.0 41.9 41.9 18:2 10.7 10.9 10.9 10.9 18:3 0.4 0.2 0.4 0.4Total SFA 66.3 58.4 45.6 45.6 Total MUFA 22.2 30.0 41.9 41.9 Total PUFA11.1 11.1 11.3 11.3 P/S 0.17 0.19 0.25 0.25 S:M:P in diet 27:9:4.624:12:4.6 19:17:4.6 19:17:4.6 PUFA % E/diet 4.6 4.6 4.6 4.6 Diet:CHO/Fat/Prot (% E) 41:41:18

The resulting lipoprotein profiles (Table 3) showed surprisingly thatthe PMF-containing diet resulted in the lowest total cholesterol (TC),VLDL, and LDL/HDL values and the highest HDL value. Owing to statisticalconfidence limits in the study, PMF in diet 711 can best be compared topalm stearin in diet 710, whereby it is concluded that TC and LDL levelswere lower for PMF compared to palm stearin. The HDL value expressed asa percentage of the total cholesterol was higher for PMF than for allother palm fractions.

TABLE 3 Body weight, blood glucose and plasma lipids of gerbils feddiets with different palm oil fractions for 3 wks Diet 710 711 713* 714Pst PMF-35 Pol IE Pol CHO:Fat:Protein (% E) 41:41:18 41:41:18 41:41:1841:41:18 Kcal/g 4.3 4.3 4.3 4.3 Body weight (g) Initial   50 ± 2   50 ±1   50 ± 2   50 ± 2 Final (after 3 wk)   59 ± 4   59 ± 4   57 ± 4   59 ±4 Gain (g/d) 0.42 ± 0.16 0.38 ± 0.18 0.28 ± 14 0.37 ± 0.14 Food Intake(g/d)  5.7 ± 0.5^(a,b,c)  5.3 ± 0.4^(a)  5.1 ± 0.2^(b)  5.2 ± 0.4^(c)(kcal/d)   25 + 1^(a,b,c)   23 ± 2^(a) 22.1 ± 1^(b)   23 ± 2^(c) F.Blood glucose at 3 wk (mg/dL)   82 ± 11^(a)   74 ± 17   69 ± 17   67 ±15^(a) Organ weights (% BW) Liver 3.26 ± 0.21 3.30 ± 0.23 3.41 ± 0.123.34 ± 0.26 Kidney 0.82 ± 0.05^(a) 0.85 ± 0.04 0.88 ± 0.04^(a) 0.86 ±0.05 Cecum 2.90 ± 0.35 2.81 ± 0.46 2.96 ± 0.30 2.86 ± 0.42 AdiposePerirenal 0.88 ± 0.36^(a) 0.68 ± 0.28 0.48 ± 0.17^(a,b) 0.73 ± 0.13^(b)Epididymal 1.39 ± 0.26^(a) 1.20 ± 0.28 1.16 ± 0.30^(a,b) 1.37 ± 0.22^(b)Total adipose 2.27 ± 0.58^(a) 1.87 ± 0.58 1.64 ± 0.31^(a,b) 2.10 ±0.32^(b) Carcass   74 ± 1^(a,b)   75 ± 1^(a)   75 ± 1^(c)   76 ± 1^(b,c)Plasma TC (mg/dL)  215 ± 50^(x)  181 ± 24^(x)  195 ± 41  201 + 47 VLDL-C(mg/dL)   54 ± 20   38 ± 10^(a)   56 ± 4   82 ± 32^(a) LDL-C (mg/dL)  88 ± 11^(x,y,z)   63 ± 18^(x)   57 ± 2^(y)   63 ± 15^(z) HDL-C (mg/dL)  98 ± 5^(a)   89 ± 6   76 ± 9   73 ± 2^(a) LDL-C/HDL-C ratio 0.90 ±0.16 0.71 ± 0.16 0.76 ± 0.11 0.85 ± 0.18 HDL-C (% of total)   41 ± 0  47 ± 4^(a)   40 ± 4   34 ± 7^(a) TG (mg/dL)   67 ± 36   56 ± 29   42 ±24   67 ± 82 Values are means ± SD (n = 8-9, except for lipoproteinsobtained by ultracentifugation of 2 or 3 samples, each representingcombined plasma from 3-4 gerbils). ^(a,b,c,d)Means in a row sharing acommon superscript are significantly different (p < 0.05) using one-wayANOVA and Fisher's PLSD test. ^(x,y,z)Means in a row sharing a commonsuperscript are significantly different (p < 0.10) using one-way ANOVAand Fisher's PLSD test.

Example 3 Gerbil Model Lipoprotein Studies with Palm Mid-FractionMargarine

Gerbil feeding experiments as described in Example 1 were used to testseveral “balanced fat” margarines containing different natural solidfats to harden vegetable oils (see Tables 4, 5 and 6). Unlike the fatblends described in Tables 1, 2 and 3, which contained high levels ofsaturated fatty acids (SFA), the margarines described herein containedfat blends providing nearly equal proportions of polyunsaturated fattyacids (PUFA) and SFA except for one margarine (B). Margarine B was usedas an experimental control to approximate the combination of fatty acidsconsumed in the typical American diet (“American Fat Blend” or AFB). Theremaining margarines E, G, and H included only natural vegetable fat andunsaturated vegetable oil blends (see Table 5) to achieve a balancedratio of PUFA and SFA (i.e., approximately a 1:1 ratio of 18:2 PUFA/SFAor “P/S” ratio). This 1:1 ratio was previously found to be beneficialfor general dietary use as described in U.S. Pat. Nos. 5,578,334,5,843,497, 6,630,192, and 7,229,653. Thus, the 18:2 PUFA:SFA “P/S”ratios for margarines E, G and H ranged from approximately 0.9 to 1.3,whereas the ratio for the AFB (margarine B) was only 0.4. Forestablishing sufficiently stable and firm textures at room temperaturewith saturated fats, margarine E incorporated 27% palm kernel oil+10%palm oil, margarine G utilized 24% palm oil and 26% palm olein, whilemargarine H utilized 15% palm mid-fraction and 8% palm oil (see Table5). With margarine H, PMF-35 contained a level of SFC of about 85% atroom temperature whereas the corresponding SFC for palm oil at roomtemperature is approximately 22%. Therefore, almost 90% of the SFC ofmargarine H is attributable to the PMF.

Table 6 provides the analytical results from these gerbil studies (dietsin Table 4) in which 41% of the dietary energy was provided by themargarines fed as the exclusive source of dietary fat. There were nostatistically significant differences in either growth or in final organor tissue weights among animals consuming the different margarines.However, the margarine B diet resulted in a very poor lipoproteinprofile with regard to standard parameters (essentially 2-fold highervalues for the parameters including TC, VLDL, LDL, LDL/HDL ratio and TG)compared to the lipoprotein profiles for all of the other diets E, G andH. Interestingly, the PMF-containing margarine H diet produced thelowest total cholesterol (TC) values. The H diet values arestatistically lower than the palm kernel oil diet values (margarine E).Otherwise, comparing each of the lipoprotein parameters among theexperimental margarines, all of the margarines tested similarly withinthe statistical confidence limits of the experiment.

TABLE 4 Gerbil diet compositions with margarines B, E, G, H Diet:719-722 Diet 719 720 721 722 Margarine B E G H CHO:Fat:Protein (% E)41:41:18 41:41:18 41:41:18 41:41:18 Kcal/g 4.3 4.3 4.3 4.3 DietIngredient (g/kg) Casein 100 100 100 100 Lactalbumin 100 100 100 100Dextrose 200 200 200 200 Cornstarch 174 + 60 (w/gel) 174 + 60 (w/gel)174 + 60 (w/gel) 174 + 60 (w/gel) Cellulose 100 100 100 100 Fat:(SFA:MUFA:PUFA % E) (17:14:7) (14:14:12) (13:13:14) (7:22:10) (P/S)(0.40) (0.90) (1.18) (1.34) AFB-Margarine B†* 250 0 0 0 Margarine E†** 0250 0 0 Margarine G†*** 0 0 250 0 Margarine H†**** 0 0 0 250 Mineral mix50 50 50 50 (Ausman - Hayes) Vitamin mix 12 12 12 12 (Hayes - Cathcart)Choline chloride 3 3 3 3 Cholesterol 0.8 0.8 0.8 0.8 Combine 60 gcornstarch with750 mL water to produce gel that is added to theremaining dry ingredients †Margarine with 80% fat and 20% water*Margarine B composition (AFB): 24% Milk Fat + 40% Tallow + 20% ChickenFat + 16% Soybean Oil **Margarine E composition: 27% Palm Kernel Oil +31% Soybean Oil + 32% Canola Oil + 10% Palm Oil ***Margarine Gcomposition (current Smart Balance): 50% Soybean Oil + 26% Palm Olein +24% Palm Oil ****Margarine H composition: 15% Palm Mid-Fraction(PMF-35) + 8% Palm Oil + 77% Canola Oil

TABLE 5 Fatty acid profile of margarine B, E, G, H Margarine MargarineMargarine B* E** G*** Margarine H**** Fatty acid % 8:0 + 10:0 0.4 1.80.0 0.0 12:0 0.8 13.0 0.1 0 14:0 4.1 4.5 0.6 0.2 16:0 22.3 11.2 25.915.3 18:0 13.2 3 4.2 2.6 18:1 35.3 34.7 32.1 54.8 18:2 14.5 25.1 32.317.5 18:3 2.0 5.1 3.9 6.8 Total SFA 41.3 33.5 30.8 18.1 Total MUFA 35.334.7 32.1 54.8 Total PUFA 16.5 30.3 32.4 24.3 P/S 0.4 0.9 1.18 1.34*Margarine B composition (AFB): 24% Milk Fat + 40% Tallow + 20% ChickenFat + 16% Soybean Oil ** Margarine E composition: 27% Palm Kernel Oil +31% Soybean Oil + 32% Canola Oil + 10% Palm Oil ***Margarine Gcomposition (current Smart Balance): 50% Soybean Oil + 26% Palm Olein +24% Palm Oil ****Margarine H composition: 15% Palm Mid-Fraction(PMF-35) + 8% Palm Oil + 77% Canola Oil

TABLE 6 Body and organ weights, blood glucose and plasma lipids ofgerbils fed diets with different fat blend margarines for 4 weeks(Gerbils Study 10) Diet 719 720 721 722 Margarine B Margarine EMargarine G Margarine H CHO:Fat:Protein (% E) 41:41:18 41:41:18 41:41:1841:41:18 kcal/g 4.3 4.3 4.3 4.3 Body weight (g) Initial   50 ± 3   50 ±3   50 ± 3   50 ± 2 Final (after 4 wk)   66 ± 6   65 ± 7   68 ± 9   65 ±8 Gain (g/d) 0.74 ± 0.22 0.70 ± 0.24 0.82 ± 0.32 0.67 ± 0.29 Food Intake(g/d)  5.8 ± 0.6  6.3 ± 0.7  6.3 ± 0.7  6.4 ± 0.9 (kcal/d)   25 ± 3   27± 3   27 ± 3   27 ± 4 F. Blood Glucose at 3 wk   80 ± 10   79 ± 10   82± 20   78 ± 9 (mg/dL) Organ weight (% BW) Liver 3.74 ± 0.46 3.68 ± 0.203.82 ± 0.29 3.86 ± 0.20 Kidney 0.88 ± 0.04 0.87 ± 0.04 0.86 ± 0.03 0.88± 0.06 Cecum 2.47 ± 0.52 2.25 ± 0.49 2.25 ± 0.57 2.43 ± 0.43 AdiposePerirenal 0.95 ± 0.31 1.02 ± 0.40 1.14 ± 0.50 0.94 ± 0.38 Epididymal1.68 ± 0.35 1.83 ± 0.59 1.91 ± 0.60 1.76 ± 0.52 Brown Fat 0.70 ± 0.160.67 ± 0.15 0.78 ± 0.23 0.67 ± 0.16 Total Adipose  3.33 + 0.78 3.52 ±1.02 3.82 ± 1.26 3.37 ± 0.88 Carcass   77 ± 1   78 ± 1   78 ± 1   78 ± 2Plasma TC (mg/dL)  200 ± 35^(a,b,c)  120 ± 25^(a,d)  103 ± 11^(b)   97 ±13^(c,d) VLDL-C (mg/dL)   53 ± 9^(,a,b,c)   29 ± 14^(a)   21 ± 5^(b)  22 ± 4^(c) LDL-C (mg/dL)   75 ± 4^(,a,b,c)   31 ± 5^(a)   28 ± 3^(b)  28 ± 2^(c) HDL-C (mg/dL)   76 ± 4^(,a,b,c)   55 ± 7^(a)   54 ± 2^(b)  47 ± 3^(c) LDL-C/HDL-C ratio 0.99 ± 0.01^(,a,b,c) 0.56 ± 0.10^(a) 0.53± 0.04^(b) 0.61 ± 0.05^(c) % HDL (of total)   37 ± 1^(,a,b,c)   48 ±2^(a)   52 ± 3^(b)   48 ± 2^(c) TG (mg/dL)  112 ± 57^(a,b)   59 ± 20^(a)  81 ± 33   62 ± 24^(b) Values are mean ± SD (n = 10) except for all thelipoprotein measurements which were obtained by ultracentrifugation of 3samples representing combined plasma from 2-4 gerbils ^(a,b,c,d..)Meansin a row sharing a common superscript are significanly different (p <0.05) using one-way ANOVA and Fisher's PLSD test

It is possible that the ability to include greater levels of PUFA intoPMF-containing margarines (cf. margarine H versus E and G) and therebyproduce margarines having higher P/S ratios may contribute to a morefavorable lipoprotein profile, in particular a low TC value. Thisability to provide higher PUFA levels relative to the SFA content ofPMF-containing margarines is attributable to the unusually high SFCmeasured at room temperature for certain PMF preparations. The ratios ofSFC relative to SFA at 20° C. was calculated for different hard fatsintended for table use. Adding a solid fat with a higher rather thanlower SFC/SFA ratio should be beneficial for hardening an unsaturatedvegetable oil while contributing a smaller amount of undesirable SFA.This SFC/SFA ratios calculated were: palm oil=0.44-0.50, palmstearin=0.99, and palm mid-fraction=1.34 for PMF-35. Therefore, certainPMF preparations are surprisingly more efficient than palm stearin athardening vegetable oils at room temperature with a minimum level ofSFA.

Between the two experiments using “low” and “higher” dietary levels ofpolyunsaturated fatty acids as linoleic acid (4.6% of dietary energy inDiets 710, 711, 713 and 714 (Example 2) and 10-14% of dietary energy inDiets 719, 720, 721 and 722 (Example 3)) the PMF palm fractionoutperformed both palm stearin and the AFB margarine with regard to oneor more of the following: lowering the plasma levels of TC, VLDL, LDL,and TG within each dietary group.

Example 4 Comparison of Fatty Acid Content of Palm Oil Fractions

The fatty acid compositions of different palm oil sources were compared,and the results are shown in Table 7. An important question is whetherany particular palm oil fractions, such as those containing saturatedfatty acid-rich triglyceride species that promote the hardening ofliquid vegetable oils, are less cholesterolemic than other saturatedfatty acid-containing triglycerides. A related question is whether anyof the three glyceride carbon positions (sn-1, sn-2 and sn-3) ispreferred with regard to improved lipoprotein metabolism for carryingsaturated fatty acids.

As shown in Table 7, each of three different palm oil-related fatscontains very different proportions of three classes of saturated fattyacid-containing triglycerides and can be evaluated for its effect onmammalian lipoprotein metabolism. The weight percentage content ofpalmitic acid is provided (“% P” based on the total fatty acidsrepresenting 100%) for the three classes of saturated fattyacid-containing triglycerides in palm stearin, PMF, and palm oil. Foreach of these fats, the relative C16:0 palmitate contents are shown inthe numerators along with the relative weights of the saturatedtriglyceride components (in denominators) for each of the fats (weightof trisaturated triglyceride (“TST”), disaturated (“DST”) andmonosaturated triglyceride (“MST”)).

TABLE 7 % P/weight TST % P/weight DST % P/weight MST Palm Stearin 52/3540/37 8/16 Palm Mid- 5/3 90/84 5/10 Fraction Palm Oil 15/9  61/46 24/35 From these numbers, it is apparent that palmitic acid is primarilyconcentrated in DST for palm mid-fraction, whereas it is distributed inTST+DST for stearin and in DST+MST for palm oil.

The numbers were obtained from the literature as follows. The case of apalm stearin with an iodine value (IV) of 30.5 is shown in Che Man etal., “Composition and Thermal Profile of Crude Palm Oil and ItsProducts,” JAOCS; 76; 237-242; 1999), where it is calculated thatapproximately 52% by weight of the palmitate content is found in TST,40% in DST, and 8% in MST. Overall, the stearin contains approximately35% by weight TST, 37% DST and 16% MST. By comparison, for a palmmid-fraction with 34.4 IV (Moran, U.S. Pat. No. 4,115,598), it iscalculated that approximately 5% by weight of the palmitate content isfound in TST, 90% in DST, and 5% in MST molecules; overall, themid-fraction contains approximately 3% by weight TST, 83% DST, 10% MST,and 4% unsaturated triglycerides. And finally, for a RBD palm oil having51.5 IV (Che Man et al., 1999), it is calculated that approximately 15%by weight of the palmitate content is found in TST, 61% in DST, and 24%in MST molecules; overall, the palm oil contains approximately 9% byweight TST, 46% DST, 35% MST, and 5% unsaturated triglycerides.

Example 5 Balancing the Ratio of Saturated Fatty Acids (SFA) andPolyunsaturated Fatty Acids (PUFA) in a PMF Fat—Canola Oil Blend

Canola oil is useful as a monounsaturated vegetable oil, for combiningwith PMF when it is desirable to limit the overall level of SFA toapproximately 20% of the fat composition, while also achieving abalanced ratio of PUFA to SFA (e.g., an approximate 1:1 ratio).Accordingly, and by way of example, a concentration of 15% by weight PMFcan be used for achieving an appropriate degree of hardening of aregular tub-type canola oil-based table spread containing 64% fat andapproximately 33% water (see Table 8). The fat blend compositioncontains approximately 80% canola oil, 15% by weight PMF-35 andapproximately 5% palm oil. The PMF coincidentally contains 64% by weightSFA contributing only about 9.6% SFA to the fat blend and 3.5% linoleicacid. With 80% (w/w) of the fat in the blend being canola oil(containing 7.5% SFA and approximately 21% linoleic acid), the canolacontributes only approximately 6.3% SFA and 16.8% linoleic acid to thecomposition. Collectively, the three fats contribute only 18.4% byweight SFA to the final blended fat composition that also contains 18%linoleic acid. Therefore, quite remarkably, this blend provides a 1:1balanced ratio of linoleic acid to SFA. Lesser or greater amounts of PMFcan be used to produce softer and harder tub spreads and even stick-typemargarines (Tables 8 and 9). Commercial preparations of 39%fat-containing light spreads have been produced in which 20% by weightPMF is used to harden a canola-soybean oil fat blend (Table 8).Commercial 78% fat-containing tub margarines have also been produced inwhich canola oil and a small amount of palm oil have been hardened with15% by weight PMF (Table 8). In addition, stick margarines have beenproduced in which a canola oil-soybean oil-palm oil blend is hardenedwith 23% by weight PMF (see Table 9). Such fat blend compositions shouldcontain little or no partially hydrogenated vegetable oil, the lattercontaining undesirable trans-fatty acids.

TABLE 8 Tub Spreads and Tub Margarine. 15% PMF 20% PMF 15% PMFDescription Regular Spread Light Spread Tub Margarine Fat Level 64% 39%78% Water 33.2150 58.1278 17.7233 Oil, Soybean 5.5000 Oil, Canola49.2800 25.6608 61.6000 Oil, Palm 5.1200 6.4000 Palm Mid Fraction-9.6000 7.8000 12.0000 Palmel 35 Oil, Olive 0.0392 Salt, Evaporated,1.6800 1.5960 1.8000 Non-Iodized Emulifiers, preservatives, 1.10501.2762 0.4767 colors, flavors, vitamins Total 100.000 100.000 100.000Total without water 66.7850 41.8722 82.2767

TABLE 9 Margarine Sticks. 23% PMF Description Marg. Sticks Fat Level 78%Water 18.2770 Oil, Soybean 6.5222 Oil, Canola 31.0445 Oil, Palm Stearin0.0000 Oil, Palm 21.9438 Palm Mid Fraction-Palmel 35 18.0000 Oil, Olive0.079 Oil, Flax seed 0.8153 Salt, Evaporated, Non-Iodized 2.2050Flavors, colors, emulsifiers 1.1132 Total 100.000 Total without water81.7230

Example 6 Formulation of Food Products

Table spreads and margarines were formulated using PMF-containingnutritional fat compositions according to the invention. Theformulations are shown in Tables 8-13. Surprisingly low levels of PMFhardstock could be used to effectively harden liquid vegetable oils.Specifically, some PMF preparations having elevated levels of solid fatcontent (SFC) as measured at or near room temperature (20-21° C. or 70°F.) showed significant advantages over palm oil and palm stearin.Elevated SFC values at room temperature were particularly useful forachieving an overall reduction in the level of fat required for additionto vegetable oils for making margarines. The use of lower levels of fatin the form of PMF contributes lesser amounts of saturated fatty acidsto unsaturated vegetable oils, e.g., to canola and soybean oils, used asthe foundation in formulating table spreads and margarines.

A 15 wt % level of PMF was found sufficient for hardening a vegetableoil blend (see, e.g., Table 8). This level is about half the level (25to 30 wt %) of PMF used by Moran in U.S. Pat. No. 4,115,598. Togetherwith the health benefit of reducing plasma cholesterol levels, thereduction in PMF usage level can provide a substantial cost savingsbecause PMF is usually priced at twice the cost per pound of palm oil.

“Nutritional Facts” for the same tub spreads, tub margarines andmargarine sticks described in Tables 8 and 9 are provided in Tables 10through Table 13 herein. These facts are routinely printed on theoutside of packaged food products sold to the consumer. In addition,these tables also provide nutritional facts for currently competitivecommercial products. Abbreviations used in these tables for otherspreads and margarines include “SB” (Smart Balance, Inc., Paramus,N.J.), “Olivio” (Olivio Premium Products, Boston, Mass.), ICBINB (‘ICan't Believe It's Not Butter,” Unilever United States), CC (CountryCrock, Unilever United States), and EB (“Earth Balance,” Smart Balance,Paramus, N.J.). In Table 10 it is apparent that the use of 15% PMFhardstock fat can produce a 64% fat-containing regular table spread witha saturated fat level (SATS) of 1.5 g per standard serving size (14 g)that is advantageously as low or lower than competing commercial spreadsincluding the 64% fat SB, the 60% fat Olivio and the 58% fat ICBINBproducts containing either chemically modified or less desirablehardening fats including interesterified fats. In Table 11, it isapparent that the use of 20% PMF hardstock fat can produce a 39%fat-containing light table spread with a saturated fat level (SATS) ofonly 1 g per standard serving size (14 g) that is advantageously lowerthan competing commercial spreads including the 39% fat SB, the 39% fatCC and the 37% fat ICBINB product. In Table 12, it is apparent that theuse of 15% PMF hardstock fat can produce a 78% fat-containing tubmargarine with a saturated fat level (SATS) of only 2 g per standardserving size (14 g) that is advantageously lower than the current EarthBalance commercial spread containing 3 g saturated fat level. In Table13, it is apparent that the use of 23% PMF hardstock fat can produce 78%fat-containing margarine sticks with a saturated fat level of only 3.5 gper standard serving size (14 g) that is advantageously as low or lowerthan competing stick margarines with 3.5-4 g saturated fats includingthe 78% fat EB stick margarine and the 78% fat ICBINB stick margarine,as well as butter with 7 g saturated fats per 80% fat in a stick.

TABLE 10 Nutritional comparison - Spreads. Nutrition Facts - SpreadsServing size 14 g 15% PMF, 64% Fat SB 64% Regular Spread Olivio 60%ICBINB 58% Calories 80 80 80 70 Calories from Fat 80 80 70 70 % DV % DV% DV % DV Total Fat 9 g 14% 9 g 14%  8 g 12%  8 g 12% Saturated Fat 2.5g 13% 1.5 g 8% 1.5 g 8 2 g 10% Trans Fat 0 g 0 g 0 g 0 g PolyunsaturatedFat 3.5 g 2 g 2 g 4 g Monounsaturated Fat 3 g 5 g 4.5 g 2 g Cholesterol0 mg  0% 0 mg 0% 0 mg 0% 0 mg  0%

TABLE 11 Nutritional comparison - Light Spreads. Nutrition Facts -Spreads Serving size 14 g 20% PMF, 39% Fat SB Light 39% Light Spread CC39% ICBINB 37% Calories 50 50 50 50 Calories from Fat 50 50 50 50 % DV %DV % DV % DV Total Fat 5 g 8% 5 g 8% 5 g 8 5 g 8% Saturated Fat 1.5 g 8%1 g 5% 1.5 g 8 1.5 g 8% Trans Fat 0 g 0 g 0 g 0 g Polyunsaturated Fat 2g 1.5 g 2.5 g 2.5 g Monounsaturated Fat 2 g 3.0 g 1 g 1 g Cholesterol 0mg 0% 0 mg 0% 0 mg 0% 0 mg 0% * Saturated fat in PMF Spreads (39% fat)is decreased to 1 g from 1.5 g (current SB 39% fat) * Lowest saturatedfat among commercial brands.

TABLE 12 Nutritional comparison - 80% Spreads. Nutrition Facts - SpreadsServing size 14 g 15% PMF, 78% Fat EB 78% Tub Margarine Calories 100 100Calories from Fat 100 100 % DV Total Fat 11 g 17% 11 g Saturated Fat 3 g15% 2 g Trans Fat 0 g 0 g Polyunsaturated Fat 2.5 g 2.5 gMonounsaturated Fat 5 g 6 g Cholesterol 0 mg 0% 0 mg

TABLE 13 Nutritional comparison Sticks. Nutrition Facts - Sticks Servingsize 14 g 23% PMF, 78% Fat Margarine ICBINB EB Sticks Sticks Butter 78%Calories 100 100 100 100 Calories from Fat 100 100 100 100 % DV % DV %DV % DV Total Fat 11 g 17% 11 g 17% 11 g 17% 11 g 17% Saturated Fat 4 g20% 3.5 g* 18% 7 g 36% 3.5 g 18% Trans Fat 0 g 0 g 0 g 0 gPolyunsaturated Fat 2.5 g 2 g 2.5 g 4.5 g Monounsaturated Fat 4.5 g 5 g4.5 g 3 g Cholesterol 0 mg  0% 0 mg  0% 30 mg 10% 0 mg  0% *Possiblewith 19.5% PMF *PMF Sticks (80% fat) are significantly superior tocurrent EB Sticks in sensory qualities *Current EB Sticks have waxymouth feel whereas PMF Sticks (80% fat) have butter-like quality*Reduced saturated fat (3 g) is sufficient for PMF Sticks containing64.5% fat

Example 7 Substitution of PMF Fat for Palm Stearin

As explained above, PMF fats are rich in DST molecules (>60% by weight),containing predominantly palmitic and/or stearic acids at the sn-1 andsn-3 positions of the molecule and oleic or linoleic acid at the sn-2position. While typical palm stearins also contain DST (˜37% by weight)they are enriched in trisaturated triglycerides or TST (˜35% by weight).It is known that different commercial preparations of PMF containvarying amounts of TST typically as tripalmitin molecules thatcontribute to the solid fat content or SFC of the fat. However TST, thatis found in abundance in palm stearin, is shown in the present studiesto be cholesterolemic, and with the PMFs selected for use herein, thelevel of both TST and MST molecules are therefore being reduced relativeto the amount of DST provided. For purposes of comparison, in theirarticle entitled “Composition and Thermal Profile of Crude Palm Oil andIts Products,” Che Man et al. (1999) report that refined, bleached anddeodorized palm oil contains, by weight, approximately 9% TST, 46% DST,35% MST, 5% tri-unsaturated triglycerides and 5% diglycerides. For thePMF selected and used herein as a hardstock, less than 5% by weight ofTST should remain in the purified PMF preparation. MST molecules, on theother hand are not particularly cholesterolemic when compared to TST,but they add undesirable SFA to the PMF preparation without contributingsignificantly to the useful solid fat content above room temperature,e.g., between approximately 22 and 30° C. Therefore, the level of MST inthe PMF should be reduced to less than 25%, and preferably to 20% orless.

TABLE 14 Physical Characteristics of Palm Oil and Palm Oil Sub-FractionsPalm Oil Palm Olein Palm Stearin Palm Mid-Fraction 35 Iodine Value50.3^(§) 56.1^(§) 30.0^(§) ^(Ø) 33-35^(§), 34.4^(#) Mettler Drop Pt. (°C.)^(§) 40.3 25.1 55.8 31-33 Fatty Acid Comp. (%)^(§) SFA 50.4 45.8 70.964.2 MUFA 39.6 42.2 23.2 32.1 PUFA 9.7 11.3  5.7  3.7 C12:0 0.3 0.3  0.2 0.1 C14:0 1.1 1.0  1.2  0.7 C16:0 44.5 40.0 64.5 57.5 C16:1 0.3 0.3 0.2  0.1 C18:0 4.3 4.3  5.0  5.6 C18:1 39.3 41.9 23.0 32.0 C18:2 9.510.9  5.3  3.5 C18:3 0.2 0.4  0.4  0.2 C20:0 0.2 0.2  0.3 Triglycerides(%) Trisaturates S₃ 8.7^(†) 2.8^(†) 35.2^(†)  3.1^(#) (1-6) DisaturatesS₂U 46.0^(†) 47.6^(†) 36.6^(†) 83.6^(#) (80-90) Monosaturates SU₂35.2^(†) 38.8^(†) 16.4^(†)  9.8^(#) (6-12) Unsaturates U₃ 5.0^(†)5.3^(†)  4.0^(†)  3.5^(#) (1-6) Solid Fat Content (%) 20° C. 20.5^(¶)1-2^(§) 70^(Ø) 86.1^(§) 25° C. 11.3^(¶) 0 60^(Ø) 74.7^(§) 30° C. 8.6^(¶)0 49^(Ø) 38.4^(§) 35° C. 2.6^(¶) 0 38Ø  0.0§ Ratio 18:1:(16:0 + 18:0)0.81 0.95  0.33  0.51 ^(Ø)Mat Sahri, et al. (2010) Palm Stearin as LowTrans Hard Stock for Margarine ^(#)Moran, U.S. Pat. No. 4,115,598^(§)Fuji Vegetable Oil Inc. sample analysis (Savannah, GA) ^(†)Che Man,et. al. (1999) Composition and Thermal Profile of Crude Palm Oil and itsProducts ^(¶)Noor Lida, et al., NMR analysis

It will be readily apparent to one skilled in the art that varyingsubstitutions and modifications may be made to the invention disclosedherein without departing from the scope and spirit of the invention. Forexample, in addition to the natural dietary fats listed herein, othersthat are not listed may be incorporated into the compositions describedherein. Likewise, other sources of disaturated triglycerides, palmiticacid, linoleic acid, and other fatty acids and fats not listed hereinthat decrease plasma levels of total cholesterol (TC) and/or LDL-Cand/or the ratio of TC/HDL-C and/or the ratio of LDL-C/HDL-C, may beincorporated into the compositions described herein, and used incombinations and concentrations not described herein, to produce naturalfat blends as well as new fats that fall within the scope of the presentinvention. Genetically engineered and naturally selected plant speciesthat produce fats whose triglycerides are structured and whose fattyacid levels are in accordance with the present invention also fallwithin the scope of the present invention. Thus, such additionalembodiments are within the scope of the present invention.

Where features or aspects of the invention are described in terms ofMarkush groups or other grouping of alternatives, the invention is alsointended to include embodiments encompassing any individual member orsubgroup of members of the Markush group or other group. Also, unlessindicated to the contrary, where various numerical values or value rangeendpoints are provided for description of certain embodiments,additional embodiments are intended as which are described by taking any2 different values as the endpoints of a range or by taking twodifferent range endpoints from specified ranges as the endpoints of anadditional range. Such ranges are also within the scope of the describedinvention. Further, specification of a numerical range including valuesgreater than one includes specific description of each integer valuewithin that range.

The invention claimed is:
 1. A margarine or table spread compositioncomprising: at least one triglyceride-based palm mid-fraction fat; andat least one triglyceride-based unsaturated vegetable oil; wherein thefat portion of the composition contains from 10% to 22% by weight ofsaid at least one palm mid-fraction fat and from 60% to 90% by weight ofsaid at least one unsaturated vegetable oil; wherein the linoleic acidcontent of the composition is from 15% to 45% by weight based on thetotal weight of fatty acids in the composition; wherein the compositionis solid or semi-solid at 20° C. and fluid at 35° C.; and wherein thecomposition is substantially free of synthetic trans-fatty acids.
 2. Thecomposition of claim 1, wherein the saturated fatty acid content of thecomposition is from 15% to 40% by weight based on the total weight offatty acids in the composition.
 3. The composition of claim 1, whereinthe saturated fatty acid content of the composition is from 15% to 23%by weight based on the total weight of fatty acids in the composition.4. The composition of claim 1, wherein the fat portion of thecomposition comprises from 75% to 90% by weight of said at least oneunsaturated vegetable oil.
 5. The composition of claim 1, wherein saidat least one unsaturated vegetable oil is selected from the groupconsisting of monounsaturated vegetable oils, polyunsaturated vegetableoils, and combinations thereof.
 6. The composition of claim 5, whereinthe at least one unsaturated vegetable oil is selected from the groupconsisting of olive oil, high oleic sunflower oil, canola oil, soybeanoil, corn oil, peanut oil, sunflower oil, safflower oil, cottonseed oil,and combinations thereof.
 7. The composition of claim 1 that issubstantially free of chemically modified vegetable oils and/or fats. 8.The composition of claim 1, wherein the palm mid-fraction fat or thevegetable oil was obtained by supplementing one or more natural fats oroils with one or more fat components isolated from a natural fat, orwith a chemically modified fat component, or with a synthetic fatcomponent.
 9. The composition of claim 1 that is substantiallycholesterol-free.
 10. The composition of claim 1 that contains from 8%to 20% by weight of disaturated triglycerides based on the total weightof triglycerides in the composition.
 11. The composition of claim 1,wherein the weight ratio of disaturated to trisaturated triglycerides isgreater than 15:1.
 12. The composition of claim 1, wherein the weightratio of disaturated to trisaturated triglycerides is greater than 20:1.13. The composition of claim 1 that contains less than 3% by weight oftrisaturated triglycerides based on the total weight of triglycerides inthe composition.
 14. The composition of claim 1 that has a solid fatcontent measured at 20° C. of from 9% to 24% by weight.
 15. Thecomposition of claim 1, wherein said at least one unsaturated vegetableoil is selected from the group consisting of monounsaturated vegetableoils, polyunsaturated vegetable oils, and combinations thereof; andwherein the ratio of linoleic acid to saturated fatty acids selectedfrom the group consisting of palmitic acid and stearic acid ranges fromabout 1:1 to about 3:1.
 16. The composition of claim 1, wherein said atleast one unsaturated vegetable oil is selected from the groupconsisting of soybean oil, canola oil, corn oil, sunflower oil,safflower oil, cottonseed oil, peanut oil, olive oil, and combinationsthereof.
 17. The composition of claim 1, wherein the weight ratio oflinoleic acid to saturated fatty acids is from about 1:1 to 3:1.
 18. Thecomposition of claim 1, wherein the trisaturated triglyceride content ofthe composition is less than 2% based on the total weight oftriglycerides in the composition.
 19. The composition of claim 1,wherein the trisaturated triglyceride content of the composition is lessthan 1% based on the total weight of triglycerides in the composition.20. The composition of claim 1 that consists essentially of said atleast one palm mid-fraction fat and said at least one unsaturatedvegetable oil.
 21. A food product comprising a fat compositioncomprising at least one triglyceride-based palm mid-fraction fat and atleast one triglyceride-based unsaturated vegetable oil; wherein the fatcomposition contains from 10% to 22% by weight of said at least one palmmid-fraction fat and from 60% to 90% by weight of said at least oneunsaturated vegetable oil; and wherein the linoleic acid content of thefat composition is from 15% to 45% by weight based on the total weightof fatty acids in the composition, wherein the food product is selectedfrom the group consisting of shortenings, baked goods, fried goods,filled dairy products, fat-containing confections, mayonnaise, and saladdressings.
 22. The food product of claim 21, wherein the fat compositionis emulsified with water.
 23. The food product of claim 22 containingfrom 40% to 85% by weight of said fat composition.
 24. The food productof claim 22 containing from 15% to 60% by weight of water.
 25. Thecomposition of claim 1, wherein the at least one palm mid-fraction fatcontains from about 60% to about 95% by weight of disaturatedtriglycerides and less than 6% by weight of trisaturated triglyceridesbased on the total triglycerides in the at least one palm mid-fractionfat, and more than 50 mol % of the disaturated triglycerides in the atleast one palm mid-fraction fat contain either palmitic acid or acombination of palmitic acid and stearic acid at the sn-1 and sn-3positions and either oleic acid or linoleic acid at the sn-2 position;wherein the disaturated triglyceride content of the composition is from8% to 23% by weight based on the total weight of triglycerides in thecomposition; wherein the weight ratio of said disaturated triglyceridesto trisaturated triglycerides in the composition is greater than 10:1;and wherein the content of (myristic acid+lauric acid) in thecomposition is less than 10% by weight based on the total weight offatty acids in the composition.
 26. The composition of claim 1, whereinthe fat portion of the composition contains from 10% to 20% by weight ofsaid at least one palm mid-fraction fat.
 27. The composition of claim 1,wherein the fat portion of the composition contains from 10% to 18% byweight of said at least one palm mid-fraction fat.
 28. The compositionof claim 1, wherein the fat portion of the composition contains 15% byweight of said at least one palm mid-fraction fat.
 29. The compositionof claim 1, wherein the linoleic acid content of the composition is from20% to 40% by weight based on the total weight of fatty acids in thecomposition.
 30. The composition of claim 1, wherein the linoleic acidcontent of the composition is from 25% to 40% by weight based on thetotal weight of fatty acids in the composition.